119 results on '"Philippe Bourin"'
Search Results
2. Functional Comparison between Healthy and Multiple Myeloma Adipose Stromal Cells
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Nicolas Espagnolle, Benjamin Hebraud, Jean-Gérard Descamps, Mélanie Gadelorge, Marie-Véronique Joubert, Laura Do Souto Ferreira, Murielle Roussel, Anne Huynh, Luc Sensébé, Louis Casteilla, Michel Attal, Hervé Avet-Loiseau, Frederic Deschaseaux, Philippe Bourin, and Jill Corre
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Internal medicine ,RC31-1245 - Abstract
Multiple myeloma (MM) is an incurable B cell neoplasia characterized by the accumulation of tumor plasma cells within the bone marrow (BM). As a consequence, bone osteolytic lesions develop in 80% of patients and remain even after complete disease remission. We and others had demonstrated that BM-derived mesenchymal stromal cells (MSCs) are abnormal in MM and thus cannot be used for autologous treatment to repair bone damage. Adipose stromal cells (ASCs) represent an interesting alternative to MSCs for cellular therapy. Thus, in this study, we wondered whether they could be a good candidate in repairing MM bone lesions. For the first time, we present a transcriptomic, phenotypic, and functional comparison of ASCs from MM patients and healthy donors (HDs) relying on their autologous MSC counterparts. In contrast to MM MSCs, MM ASCs did not exhibit major abnormalities. However, the changes observed in MM ASCs and the supportive property of ASCs on MM cells question their putative and safety uses at an autologous or allogenic level.
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- 2020
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3. Human adipose stromal cells (ASC) for the regeneration of injured cartilage display genetic stability after in vitro culture expansion.
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Simona Neri, Philippe Bourin, Julie-Anne Peyrafitte, Luca Cattini, Andrea Facchini, and Erminia Mariani
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Medicine ,Science - Abstract
Mesenchymal stromal cells are emerging as an extremely promising therapeutic agent for tissue regeneration due to their multi-potency, immune-modulation and secretome activities, but safety remains one of the main concerns, particularly when in vitro manipulation, such as cell expansion, is performed before clinical application. Indeed, it is well documented that in vitro expansion reduces replicative potential and some multi-potency and promotes cell senescence. Furthermore, during in vitro aging there is a decrease in DNA synthesis and repair efficiency thus leading to DNA damage accumulation and possibly inducing genomic instability. The European Research Project ADIPOA aims at validating an innovative cell-based therapy where autologous adipose stromal cells (ASCs) are injected in the diseased articulation to activate regeneration of the cartilage. The primary objective of this paper was to assess the safety of cultured ASCs. The maintenance of genetic integrity was evaluated during in vitro culture by karyotype and microsatellite instability analysis. In addition, RT-PCR array-based evaluation of the expression of genes related to DNA damage signaling pathways was performed. Finally, the senescence and replicative potential of cultured cells was evaluated by telomere length and telomerase activity assessment, whereas anchorage-independent clone development was tested in vitro by soft agar growth. We found that cultured ASCs do not show genetic alterations and replicative senescence during the period of observation, nor anchorage-independent growth, supporting an argument for the safety of ASCs for clinical use.
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- 2013
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4. Adult stromal cells derived from human adipose tissue provoke pancreatic cancer cell death both in vitro and in vivo.
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Beatrice Cousin, Emmanuel Ravet, Sandrine Poglio, Fabienne De Toni, Mélanie Bertuzzi, Hubert Lulka, Ismahane Touil, Mireille André, Jean-Louis Grolleau, Jean-Marie Péron, Jean-Pierre Chavoin, Philippe Bourin, Luc Pénicaud, Louis Casteilla, Louis Buscail, and Pierre Cordelier
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Medicine ,Science - Abstract
BackgroundNormal tissue homeostasis is maintained by dynamic interactions between epithelial cells and their microenvironment. Disrupting this homeostasis can induce aberrant cell proliferation, adhesion, function and migration that might promote malignant behavior. Indeed, aberrant stromal-epithelial interactions contribute to pancreatic ductal adenocarcinoma (PDAC) spread and metastasis, and this raises the possibility that novel stroma-targeted therapies represent additional approaches for combating this malignant disease. The aim of the present study was to determine the effect of human stromal cells derived from adipose tissue (ADSC) on pancreatic tumor cell proliferation.Principal findingsCo-culturing pancreatic tumor cells with ADSC and ADSC-conditioned medium sampled from different donors inhibited cancer cell viability and proliferation. ADSC-mediated inhibitory effect was further extended to other epithelial cancer-derived cell lines (liver, colon, prostate). ADSC conditioned medium induced cancer cell necrosis following G1-phase arrest, without evidence of apoptosis. In vivo, a single intra-tumoral injection of ADSC in a model of pancreatic adenocarcinoma induced a strong and long-lasting inhibition of tumor growth.ConclusionThese data indicate that ADSC strongly inhibit PDAC proliferation, both in vitro and in vivo and induce tumor cell death by altering cell cycle progression. Therefore, ADSC may constitute a potential cell-based therapeutic alternative for the treatment of PDAC for which no effective cure is available.
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- 2009
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5. Supplementary Figure 3 from Bioactivity and Prognostic Significance of Growth Differentiation Factor GDF15 Secreted by Bone Marrow Mesenchymal Stem Cells in Multiple Myeloma
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Philippe Bourin, Michel Attal, Jean-Jacques Fournié, Thierry Facon, Philippe Moreau, Bernard Klein, Pierre Cordelier, Mélanie Gadelorge, Anne Huynh, Murielle Roussel, Hervé Avet-Loiseau, Benjamin Hébraud, Nicolas Espagnolle, Elodie Labat, and Jill Corre
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PDF file - 84K, Baseline phosphorylation of pT308 Akt in MOLP-6 and MM1.S cells
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- 2023
6. Supplementary Figure 1 from Bioactivity and Prognostic Significance of Growth Differentiation Factor GDF15 Secreted by Bone Marrow Mesenchymal Stem Cells in Multiple Myeloma
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Philippe Bourin, Michel Attal, Jean-Jacques Fournié, Thierry Facon, Philippe Moreau, Bernard Klein, Pierre Cordelier, Mélanie Gadelorge, Anne Huynh, Murielle Roussel, Hervé Avet-Loiseau, Benjamin Hébraud, Nicolas Espagnolle, Elodie Labat, and Jill Corre
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PDF file - 67K, Effect of GDF15 on T308 Akt phosphorylation of MOLP-6 stroma-dependent cells in culture condition with 10% fetal calf serum
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- 2023
7. Supplementary Figure 4 from Bioactivity and Prognostic Significance of Growth Differentiation Factor GDF15 Secreted by Bone Marrow Mesenchymal Stem Cells in Multiple Myeloma
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Philippe Bourin, Michel Attal, Jean-Jacques Fournié, Thierry Facon, Philippe Moreau, Bernard Klein, Pierre Cordelier, Mélanie Gadelorge, Anne Huynh, Murielle Roussel, Hervé Avet-Loiseau, Benjamin Hébraud, Nicolas Espagnolle, Elodie Labat, and Jill Corre
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PDF file - 90K, Effect of GDF15 on ERK1/2 phosphorylation in MOLP-6 and MM1.S cells
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- 2023
8. Supplementary Figure 5 from Bioactivity and Prognostic Significance of Growth Differentiation Factor GDF15 Secreted by Bone Marrow Mesenchymal Stem Cells in Multiple Myeloma
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Philippe Bourin, Michel Attal, Jean-Jacques Fournié, Thierry Facon, Philippe Moreau, Bernard Klein, Pierre Cordelier, Mélanie Gadelorge, Anne Huynh, Murielle Roussel, Hervé Avet-Loiseau, Benjamin Hébraud, Nicolas Espagnolle, Elodie Labat, and Jill Corre
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PDF file - 44K, Concentration of GDF15 measured by ELISA in 72 hours culture supernatants of primary BM-MSCs and MM cells from 3 patients, and MM cell lines MOLP-6 and MM1.S
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- 2023
9. Supplementary Figure 6 from Bioactivity and Prognostic Significance of Growth Differentiation Factor GDF15 Secreted by Bone Marrow Mesenchymal Stem Cells in Multiple Myeloma
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Philippe Bourin, Michel Attal, Jean-Jacques Fournié, Thierry Facon, Philippe Moreau, Bernard Klein, Pierre Cordelier, Mélanie Gadelorge, Anne Huynh, Murielle Roussel, Hervé Avet-Loiseau, Benjamin Hébraud, Nicolas Espagnolle, Elodie Labat, and Jill Corre
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PDF file - 43K, Correlation between the concentrations of GDF15 measured by ELISA in bone marrow (BM) plasma and in blood plasma in 24 patients with newly diagnosed MM
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- 2023
10. Supplementary Figure 8 from Bioactivity and Prognostic Significance of Growth Differentiation Factor GDF15 Secreted by Bone Marrow Mesenchymal Stem Cells in Multiple Myeloma
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Philippe Bourin, Michel Attal, Jean-Jacques Fournié, Thierry Facon, Philippe Moreau, Bernard Klein, Pierre Cordelier, Mélanie Gadelorge, Anne Huynh, Murielle Roussel, Hervé Avet-Loiseau, Benjamin Hébraud, Nicolas Espagnolle, Elodie Labat, and Jill Corre
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PDF file - 67K, Transfection of BM-MSC with plasmids encoding shRNA targeting GDF15 and co-culture with MOLP-6 cells
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- 2023
11. Toll-like receptor 4 selective inhibition in medullar microenvironment alters multiple myeloma cell growth
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Arthur Justo, Hervé Avet-Loiseau, Ludovic Martinet, Malik Hamaidia, Jill Corre, Jean-Gerard Descamps, Frédéric Deschaseaux, Laura Do Souto Ferreira, Melanie Gadelorge, Philippe Bourin, Nicolas Espagnolle, Lea Lemaitre, Marie Veronique Joubert, and Nicolas Reina
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Cell ,Mice ,In vivo ,medicine ,Tumor Microenvironment ,Animals ,Receptor ,Interleukin 6 ,Cells, Cultured ,Toll-like receptor ,biology ,Chemistry ,Cell growth ,Interleukin-6 ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Hematology ,Stimulus Report ,Toll-Like Receptor 4 ,medicine.anatomical_structure ,Cancer research ,TLR4 ,biology.protein ,Multiple Myeloma - Abstract
Key Points TLR4 is overexpressed on MM MSCs, and its activation promotes the crosstalk with MM cells in enhancing protumoral factors.Specifically targeting TLR4 on MM MSCs strengthens conventional MM therapy and weakens MM development in vivo., Visual Abstract, Bone marrow (BM) mesenchymal stromal cells (MSCs) are abnormal in multiple myeloma (MM) and play a critical role by promoting growth, survival, and drug resistance of MM cells. We observed higher Toll-like receptor 4 (TLR4) gene expression in MM MSCs than in MSCs from healthy donors. At the clinical level, we highlighted that TLR4 expression in MM MSCs evolves in parallel with the disease stage. Thus, we reasoned that the TLR4 axis is pivotal in MM by increasing the protumor activity of MSCs. Challenging primary MSCs with TLR4 agonists increased the expression of CD54 and interleukin-6 (IL-6), 2 factors directly implicated in MM MSC-MM cell crosstalk. Then, we evaluated the therapeutic efficacy of a TLR4 antagonist combined or not with conventional treatment in vitro with MSC-MM cell coculture and in vivo with the Vk*MYC mouse model. Selective inhibition of TLR4 specifically reduced the MM MSC ability to support the growth of MM cells in an IL-6-dependent manner and delayed the development of MM in the Vk*MYC mouse model by altering the early disease phase in vivo. For the first time, we demonstrate that specific targeting of the pathological BM microenvironment via TLR4 signaling could be an innovative approach to alter MM pathology development.
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- 2022
12. Integrated Transcriptomic, Phenotypic, and Functional Study Reveals Tissue-Specific Immune Properties of Mesenchymal Stromal Cells
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Philippe Collas, Nadège Bescher, Erwan Flecher, Karin Tarte, Joelle Dulong, Benjamin Hébraud, David Roulois, Louis Casteilla, Céline Monvoisin, Léa Verdière, Mélanie Gadelorge, Céline Pangault, Cédric Ménard, Vonick Sibut, Luc Sensebé, Philippe Bourin, Nicolas Espagnolle, Nicolas Bertheuil, Isabelle Bezier, Role des Cellules Dendritiques Dans la Regulation des Effecteurs de l'Immunite Antitumorale, Université Paris-Sud - Paris 11 (UP11)-Institut National de la Santé et de la Recherche Médicale (INSERM), Etablissement français du sang [Rennes] (EFS Bretagne), Centre de Recherche en Cancérologie / Nantes - Angers (CRCNA), Centre hospitalier universitaire de Nantes (CHU Nantes)-Faculté de Médecine d'Angers-Centre Hospitalier Universitaire d'Angers (CHU Angers), PRES Université Nantes Angers Le Mans (UNAM)-PRES Université Nantes Angers Le Mans (UNAM)-Centre National de la Recherche Scientifique (CNRS)-Hôpital Laennec-Institut National de la Santé et de la Recherche Médicale (INSERM)-Hôtel-Dieu de Nantes, Institut Universitaire du Cancer de Toulouse - Oncopole (IUCT Oncopole - UMR 1037), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-CHU Toulouse [Toulouse]-Institut National de la Santé et de la Recherche Médicale (INSERM), Microenvironment, Cell Differentiation, Immunology and Cancer (MICMAC), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Microenvironnement et cancer (MiCa), Recherches Avicoles (SRA), Institut National de la Recherche Agronomique (INRA), Laboratoire de Thérapie Cellulaire, EFS, Service de chirurgie thoracique cardiaque et vasculaire [Rennes], Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Hôpital Pontchaillou-CHU Pontchaillou [Rennes], Institut d'Electronique et de Télécommunications de Rennes (IETR), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)-SUPELEC-Centre National de la Recherche Scientifique (CNRS), STROMALab, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS), Akershus University Hospital [Lørenskog], Laboratoire de thérapie cellulaire (EFS), Etablissement Français du Sang, ANR-10-IBHU_0005, Agence Nationale de la Recherche, APR 2016, Etablissement Français du Sang, INCa PLBIO-17-219, Institut National Du Cancer, Appel d'offre local 2008, CHU Toulouse, Service de chirurgie thoracique cardiaque et vasculaire [Rennes] = Thoracic and Cardiovascular Surgery [Rennes], CHU Pontchaillou [Rennes], Centre National de la Recherche Scientifique (CNRS)-Ecole Supérieure d'Electricité - SUPELEC (FRANCE)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Institut National des Sciences Appliquées (INSA)-Université de Rennes (UNIV-RENNES)-Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES), Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Etablissement Français du Sang-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université de Toulouse (UT)-Université de Toulouse (UT)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Rennes (UR)-Institut National des Sciences Appliquées - Rennes (INSA Rennes), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Ecole Supérieure d'Electricité - SUPELEC (FRANCE)-Centre National de la Recherche Scientifique (CNRS), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS), ANR-10-IBHU-0005,CESTI (TSI-IHU),Centre Européen des Sciences de la Transplantation et de l'Immunothérapie (TSI-IHU)(2010), Ménard, Cédric, Instituts Hospitalo-Universitaires B - Centre Européen des Sciences de la Transplantation et de l'Immunothérapie (TSI-IHU) - - CESTI (TSI-IHU)2010 - ANR-10-IBHU-0005 - IBHU - VALID, and Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Institut National de la Santé et de la Recherche Médicale (INSERM)
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Male ,0301 basic medicine ,Karin ,UMR U1236 MICMAC ,Cell ,Philippe ,Adipose tissue ,0302 clinical medicine ,Gene expression ,Cells, Cultured ,ComputingMilieux_MISCELLANEOUS ,INSERM ,Bone marrow stromal cells (BMSCs) ,Etablissement Français du Sang ,CSA21 ,Cell Differentiation ,Middle Aged ,Phenotype ,Immunogenicity ,Cell interactions Journal Section: Translational and Clinical Research Cell Types: Bone Marrow Stem Cells ,Cell biology ,medicine.anatomical_structure ,CHU Rennes Keywords: adipose stem cells ,Molecular Medicine ,[SDV.IMM]Life Sciences [q-bio]/Immunology ,Female ,STROMALAB Tarte ,Adult ,Stromal cell ,Nicolas ,[SDV.IMM] Life Sciences [q-bio]/Immunology ,Adipose Stem Cells/VSF ,Biology ,Young Adult ,03 medical and health sciences ,Immune system ,medicine ,Humans ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Cell Biology ,Direction Espagnolle ,Université Rennes 1 ,030104 developmental biology ,SITI Laboratory ,Bone marrow ,Transcriptome ,030217 neurology & neurosurgery ,Immunosuppression ,Developmental Biology - Abstract
Clinical-grade mesenchymal stromal cells (MSCs) can be expanded from bone marrow and adipose tissue to treat inflammatory diseases and degenerative disorders. However, the influence of their tissue of origin on their functional properties, including their immunosuppressive activity, remains unsolved. In this study, we produced paired bone marrow-derived mesenchymal stromal cell (BM-MSC) and adipose-derived stromal cell (ASC) batches from 14 healthy donors. We then compared them using transcriptomic, phenotypic, and functional analyses and validated our results on purified native MSCs to infer which differences were really endowed by tissue of origin. Cultured MSCs segregated together owing to their tissue of origin based on their gene expression profile analyzed using differential expression and weighted gene coexpression network analysis. This translated into distinct immune-related gene signatures, phenotypes, and functional cell interactions. Importantly, sorted native BM-MSCs and ASCs essentially displayed the same distinctive patterns than their in vitro-expanded counterparts. As a whole, ASCs exhibited an immune profile consistent with a stronger inhibition of immune response and a lower immunogenicity, supporting the use of adipose tissue as a valuable source for clinical applications.
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- 2020
13. Functional Comparison between Healthy and Multiple Myeloma Adipose Stromal Cells
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Marie-Véronique Joubert, Laura Do Souto Ferreira, Jean-Gérard. Descamps, Frédéric Deschaseaux, Hervé Avet-Loiseau, Philippe Bourin, Murielle Roussel, Luc Sensebé, Louis Casteilla, Benjamin Hebraud, Anne Huynh, Nicolas Espagnolle, Mélanie Gadelorge, Michel Attal, and Jill Corre
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0301 basic medicine ,Pathology ,medicine.medical_specialty ,Stromal cell ,Article Subject ,Adipose tissue ,Cell therapy ,Transcriptome ,03 medical and health sciences ,0302 clinical medicine ,medicine ,Molecular Biology ,Internal medicine ,Multiple myeloma ,B cell ,business.industry ,Mesenchymal stem cell ,Cell Biology ,medicine.disease ,RC31-1245 ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Bone marrow ,business ,Research Article - Abstract
Multiple myeloma (MM) is an incurable B cell neoplasia characterized by the accumulation of tumor plasma cells within the bone marrow (BM). As a consequence, bone osteolytic lesions develop in 80% of patients and remain even after complete disease remission. We and others had demonstrated that BM-derived mesenchymal stromal cells (MSCs) are abnormal in MM and thus cannot be used for autologous treatment to repair bone damage. Adipose stromal cells (ASCs) represent an interesting alternative to MSCs for cellular therapy. Thus, in this study, we wondered whether they could be a good candidate in repairing MM bone lesions. For the first time, we present a transcriptomic, phenotypic, and functional comparison of ASCs from MM patients and healthy donors (HDs) relying on their autologous MSC counterparts. In contrast to MM MSCs, MM ASCs did not exhibit major abnormalities. However, the changes observed in MM ASCs and the supportive property of ASCs on MM cells question their putative and safety uses at an autologous or allogenic level.
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- 2019
14. Elaboration and evaluation of alginate foam scaffolds for soft tissue engineering
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Benjamin Duployer, Philippe Bourin, Caroline Ceccaldi, Brigitte Sallerin, Angelo Parini, Christophe Tenailleau, Daniel Cussac, Sophie Girod Fullana, Raya Bushkalova, Centre interuniversitaire de recherche et d'ingenierie des matériaux (CIRIMAT), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut de Chimie du CNRS (INC), Institut des Maladies Métaboliques et Cardiovasculaires (I2MC), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Université Fédérale Toulouse Midi-Pyrénées, EFS ALPES MEDITERRANEE, CHU Toulouse [Toulouse], Centre National de la Recherche Scientifique - CNRS (FRANCE), Etablissement Français du Sang - EFS Midi-Pyrénées (FRANCE), Institut National Polytechnique de Toulouse - Toulouse INP (FRANCE), Institut National de la Santé et de la Recherche Médicale - INSERM (FRANCE), Université Toulouse III - Paul Sabatier - UT3 (FRANCE), and Centre Hospitalier Universitaire de Toulouse - CHU Toulouse (FRANCE)
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0301 basic medicine ,Pore size ,Scaffold ,Materials science ,Alginates ,Matériaux ,Surfactants ,Foam-based scaffolds ,Pharmaceutical Science ,Paracrine activity ,Nanotechnology ,Biocompatible Materials ,02 engineering and technology ,03 medical and health sciences ,Glucuronic Acid ,Soft tissue engineering ,Humans ,Polymer scaffold ,[SDV.IB.BIO]Life Sciences [q-bio]/Bioengineering/Biomaterials ,Tissue Engineering ,Tissue Scaffolds ,Hexuronic Acids ,Mesenchymal stem cell ,Alginate ,021001 nanoscience & nanotechnology ,Biocompatible material ,Porous scaffold ,030104 developmental biology ,[CHIM.POLY]Chemical Sciences/Polymers ,Mesenchymal stem cells ,0210 nano-technology ,Porosity - Abstract
International audience; Controlling microarchitecture in polymer scaffolds is a priority in material design for soft tissue applications. This paper reports for the first time the elaboration of alginate foam-based scaffolds for mesenchymal stem cell (MSC) delivery and a comparative study of various surfactants on the final device performance. The use of surfactants permitted to obtain highly interconnected porous scaffolds with tunable pore size on surface and in cross-section. Their mechanical properties in compression appeared to be adapted to soft tissue engineering. Scaffold structures could sustain MSC proliferation over 14 days. Paracrine activity of scaffold-seeded MSCs varied with the scaffold structure and growth factors release was globally improved in comparison with control alginate scaffolds. Our results provide evidence that exploiting different surfactant types for alginate foam preparation could be an original method to obtain biocompatible scaffolds with tunable architecture for soft tissue engineering.
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- 2017
15. Phase I trial: the use of autologous cultured adipose-derived stroma/stem cells to treat patients with non-revascularizable critical limb ischemia
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Sophie Dupuis-Coronas, Bertrand Leobon, Jean-Sébastien Silvestre, Philippe Bourin, Louis Casteilla, Mélanie Gadelorge, Bertrand Saint-Lebese, Marion Taurand, Sandrine Fleury, Jean-Louis Grolleau, Valérie Planat-Benard, Fabian Gross, Julie-Anne Peyrafitte, and Alessandra Bura
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Adult ,Male ,Cancer Research ,medicine.medical_specialty ,medicine.medical_treatment ,Immunology ,Cell Culture Techniques ,Neovascularization, Physiologic ,Adipose tissue ,Revascularization ,Injections, Intramuscular ,Cell therapy ,Peripheral Arterial Disease ,Ischemia ,medicine ,Humans ,Immunology and Allergy ,Cells, Cultured ,Genetics (clinical) ,Aged ,Aged, 80 and over ,Homeodomain Proteins ,Transplantation ,business.industry ,Mesenchymal stem cell ,Extremities ,Nanog Homeobox Protein ,Cell Biology ,Critical limb ischemia ,Middle Aged ,Surgery ,Adult Stem Cells ,Treatment Outcome ,Adipose Tissue ,Oncology ,Feasibility Studies ,Female ,Stromal Cells ,medicine.symptom ,Stem cell ,Wound healing ,business ,Octamer Transcription Factor-3 ,Stem Cell Transplantation - Abstract
Non-revascularizable critical limb ischemia (CLI) is the most severe stage of peripheral arterial disease, with no therapeutic option. Extensive preclinical studies have demonstrated that adipose-derived stroma cell (ASC) transplantation strongly improves revascularization and tissue perfusion in ischemic limbs. This study, named ACellDREAM, is the first phase I trial to evaluate the feasibility and safety of intramuscular injections of autologous ASC in non-revascularizable CLI patients.Seven patients were consecutively enrolled, on the basis of the following criteria: (i) lower-limb rest pain or ulcer; (ii) ankle systolic oxygen pressure 50 or 70 mm Hg for non-diabetic and diabetic patients, respectively, or first-toe systolic oxygen pressure 30 mm Hg or 50 mm Hg for non-diabetic and diabetic patients, respectively; (iii) not suitable for revascularization. ASCs from abdominal fat were grown for 2 weeks and were then characterized.More than 200 million cells were obtained, with almost total homogeneity and no karyotype abnormality. The expressions of stemness markers Oct4 and Nanog were very low, whereas expression of telomerase was undetectable in human ASCs compared with human embryonic stem cells. ASCs (10(8)) were then intramuscularly injected into the ischemic leg of patients, with no complication, as judged by an independent committee. Trans-cutaneous oxygen pressure tended to increase in most patients. Ulcer evolution and wound healing showed improvement.These data demonstrate the feasibility and safety of autologous ASC transplantation in patients with objectively proven CLI not suitable for revascularization. The improved wound healing also supports a putative functional efficiency.
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- 2014
16. <scp>CD</scp> 146 expression on mesenchymal stem cells is associated with their vascular smooth muscle commitment
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Frédéric Deschaseaux, Nicolas Espagnolle, Luc Sensebé, Mélanie Gadelorge, Philippe Bourin, and Fabien Guilloton
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Vascular smooth muscle ,proliferation ,Cellular differentiation ,Molecular Sequence Data ,Myocytes, Smooth Muscle ,Population ,CD146 Antigen ,Cell Separation ,Biology ,Muscle, Smooth, Vascular ,Transforming Growth Factor beta1 ,Transcriptome ,medicine ,Humans ,education ,Cells, Cultured ,Cell Proliferation ,mesenchymal stem cells ,education.field_of_study ,Mesenchymal stem cell ,Cell Differentiation ,Original Articles ,differentiation ,Cell Biology ,Cell biology ,Phenotype ,medicine.anatomical_structure ,CD146 ,Adipogenesis ,Molecular Medicine ,Fibroblast Growth Factor 2 ,Bone marrow ,vascular smooth muscle cell - Abstract
Bone marrow mesenchymal stem cells (MSCs) are plastic adherent cells that can differentiate into various tissue lineages, including osteoblasts, adipocytes and chondrocytes. However, this progenitor property is not shared by all cells within the MSC population. In addition, MSCs vary in their proliferation capacity and expression of markers. Because of heterogeneity of CD146 expression in the MSC population, we compared CD146(-/Low) and CD146(High) cells under clonal conditions and after sorting of the non-clonal cell population to determine whether this expression is associated with specific functions. CD146(-/Low) and CD146(High) bone marrow MSCs did not differ in colony-forming unit-fibroblast number, osteogenic, adipogenic and chondrogenic differentiation or in vitro haematopoietic-supportive activity. However, CD146(-/Low) clones proliferated slightly but significantly faster than did CD146(High) clones. In addition, a strong expression of CD146 molecule was associated with a commitment to a vascular smooth muscle cell (VSMC) lineage characterized by a strong up-regulation of calponin-1 and SM22α expression and an ability to contract collagen matrix. Thus, within a bone marrow MSC population, certain subpopulations characterized by high expression of CD146, are committed towards a VSMC lineage.
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- 2013
17. Adipose mesenchymal stem cells protect chondrocytes from degeneration associated with osteoarthritis
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Cristina Manferdini, Elena Gabusi, Philippe Bourin, Marie Maumus, Rosanna Ferreira, Gina Lisignoli, Andrea Facchini, Danièle Noël, Julie-Anne Peyrafitte, Karine Toupet, and Christian Jorgensen
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Adult ,Male ,Adipose tissue ,Apoptosis ,Biology ,03 medical and health sciences ,Chondrocytes ,0302 clinical medicine ,Osteoarthritis ,Adipocytes ,medicine ,Humans ,Secretion ,Cells, Cultured ,030304 developmental biology ,Medicine(all) ,0303 health sciences ,Mesenchymal stem cell ,Cell Differentiation ,Mesenchymal Stem Cells ,Cell Biology ,General Medicine ,Middle Aged ,Phenotype ,Cell biology ,medicine.anatomical_structure ,Cell culture ,Immunology ,Female ,Bone marrow ,Stem cell ,030217 neurology & neurosurgery ,Developmental Biology - Abstract
Our work aimed at evaluating the role of adipose stem cells (ASC) on chondrocytes from osteoarthritic (OA) patients and identifying the mediators involved. We used primary chondrocytes, ASCs from different sources and bone marrow mesenchymal stromal cells (MSC) from OA donors. ASCs or MSCs were co-cultured with chondrocytes in a minimal medium and using cell culture inserts. Under these conditions, ASCs did not affect the proliferation of chondrocytes but significantly decreased camptothecin-induced apoptosis. Both MSCs and ASCs from different sources allowed chondrocytes in the cocultures maintaining a stable expression of markers specific for a mature phenotype, while expression of hypertrophic and fibrotic markers was decreased. A number of factors known to regulate the chondrocyte phenotype (IL-1β, IL-1RA, TNF-α) and matrix remodeling (TIMP-1 and -2, MMP-1 and -9, TSP-1) were not affected. However, a significant decrease of TGF-β1 secretion by chondrocytes and induction of HGF secretion by ASCs was observed. Addition of a neutralizing anti-HGF antibody reversed the anti-fibrotic effect of ASCs whereas hypertrophic markers were not modulated. In summary, ASCs are an interesting source of stem cells for efficiently reducing hypertrophy and dedifferentiation of chondrocytes, at least partly via the secretion of HGF. This supports the interest of using these cells in therapies for osteo-articular diseases.
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- 2013
18. Long-Term Detection of Human Adipose-Derived Mesenchymal Stem Cells After Intraarticular Injection in SCID Mice
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Nelly Pirot, Rosanna Ferreira, Peter L. E. M. van Lent, Christian Jorgensen, Julie-Anne Peyrafitte, Danièle Noël, Louis Casteilla, Béatrice Orsetti, Karine Toupet, Philippe Bourin, and Marie Maumus
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0303 health sciences ,Biodistribution ,Pathology ,medicine.medical_specialty ,business.industry ,Immunology ,Mesenchymal stem cell ,Adipose tissue ,3. Good health ,Cell therapy ,03 medical and health sciences ,0302 clinical medicine ,medicine.anatomical_structure ,Rheumatology ,030220 oncology & carcinogenesis ,Toxicity ,medicine ,Immunology and Allergy ,Distribution (pharmacology) ,Pharmacology (medical) ,Bone marrow ,Stem cell ,business ,030304 developmental biology - Abstract
Objective Mesenchymal stem cells (MSCs) represent a promising tool for cell therapy for several disorders, among them the osteoarticular diseases. For such clinical applications, intraarticular (IA) injection of MSCs may be favored for higher levels of safety and targeting of specific joints. Although the safety of intravenous (IV) administration of MSCs has been reported in a number of clinical trials, the safety and biodistribution of MSCs after IA injection have not been tested. Our objective was to assess the toxicity of clinical-grade human adipose-derived MSCs (AD-MSCs), as well as their biodistribution, after IA injection into SCID mice. Methods SCID mice received IA or IV administration of 106 human AD-MSCs. Several tissues were recovered at different time points and processed for histologic assessment or real-time polymerase chain reaction (PCR) analysis. A highly sensitive assay was used to monitor the distribution of AD-MSCs, based on amplification of human-specific Alu sequences. Results Absence of toxicity was observed after AD-MSC infusion. Alu PCR assay revealed a high sensitivity (1 human AD-MSC/105 murine cells), with a large linear range (1–5 × 104/105 murine cells). Importantly, 15% of the IA-injected AD-MSCs were detectable in the joint for the first month and 1.5% of the AD-MSCs engrafted over the long term, at least 6 months. AD-MSCs were observed in the injected joints and in areas of tissue referred to as stem cell niches, such as the bone marrow, adipose tissue, and muscle. Conclusion These data support the feasibility and safety of using IA delivery of human AD-MSCs in the treatment of rheumatic diseases that affect the joints.
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- 2013
19. Stromal cells from the adipose tissue-derived stromal vascular fraction and culture expanded adipose tissue-derived stromal/stem cells: a joint statement of the International Federation for Adipose Therapeutics and Science (IFATS) and the International Society for Cellular Therapy (ISCT)
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Heinz Redl, Jeffrey M. Gimble, Keith L. March, J. Peter Rubin, Louis Casteilla, Bruce A. Bunnell, Adam J. Katz, Kotaro Yoshimura, Philippe Bourin, and Massimo Dominici
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Cancer Research ,Pathology ,Cell- and Tissue-Based Therapy ,Adipose tissue ,Antigens, CD34 ,Regenerative Medicine ,Adipose-derived Stromal Vascular Fraction Cells ,Adipocytes ,Immunology and Allergy ,characterization ,Cells, Cultured ,Genetics (clinical) ,education.field_of_study ,Cultured ,Mesenchymal Stromal Cells ,Stem Cells ,Cell Differentiation ,Reference Standards ,Stromal vascular fraction ,Flow Cytometry ,adipose tissue ,Adipose Tissue ,Oncology ,Molecular Medicine ,Stem cell ,medicine.medical_specialty ,adipose-derived stromal/stem cells ,Stromal cell ,phenotype ,Cells ,Immunology ,Population ,Article ,medicine ,Humans ,CD90 ,Obesity ,Antigens ,education ,function ,Transplantation ,business.industry ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Cell Biology ,stromal vascular fraction ,Stromal Cells ,CD34 ,business - Abstract
Background aims Adipose tissue is a rich and very convenient source of cells for regenerative medicine therapeutic approaches. However, a characterization of the population of adipose-derived stromal and stem cells (ASCs) with the greatest therapeutic potential remains unclear. Under the authority of International Federation of Adipose Therapeutics and International Society for Cellular Therapy, this paper sets out to establish minimal definitions of stromal cells both as uncultured stromal vascular fraction (SVF) and as an adherent stromal/stem cells population. Methods Phenotypic and functional criteria for the identification of adipose-derived cells were drawn from the literature. Results In the SVF, cells are identified phenotypically by the following markers: CD45-CD235a-CD31-CD34+. Added value may be provided by both a viability marker and the following surface antigens: CD13, CD73, CD90 and CD105. The fibroblastoid colony-forming unit assay permits the evaluation of progenitor frequency in the SVF population. In culture, ASCs retain markers in common with other mesenchymal stromal/stem cells (MSCs), including CD90, CD73, CD105, and CD44 and remain negative for CD45 and CD31. They can be distinguished from bone-marrow-derived MSCs by their positivity for CD36 and negativity for CD106. The CFU-F assay is recommended to calculate population doublings capacity of ASCs. The adipocytic, chondroblastic and osteoblastic differentiation assays serve to complete the cell identification and potency assessment in conjunction with a quantitative evaluation of the differentiation either biochemically or by reverse transcription polymerase chain reaction. Conclusions The goal of this paper is to provide initial guidance for the scientific community working with adipose-derived cells and to facilitate development of international standards based on reproducible parameters.
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- 2013
20. Adipose-Derived Mesenchymal Stem Cells Exert Antiinflammatory Effects on Chondrocytes and Synoviocytes From Osteoarthritis Patients Through Prostaglandin E-2
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Christian Jorgensen, Cristina Manferdini, Giuseppe Filardo, Philippe Bourin, Marie Maumus, Anna Piacentini, Andrea Facchini, Gina Lisignoli, Sandrine Fleury-Cappellesso, Julie-Anne Peyrafitte, Danièle Noël, Elena Gabusi, C. Manferdini, M. Maumu, E. Gabusi, A. Piacentini, G. Filardo, J. Peyrafitte, C. Jorgensen, P. Bourin, S. Fleury-Cappellesso, A. Facchini, D. Noël, and G. Lisignoli
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Cartilage, Articular ,Male ,Chemokine ,ANABOLIC CYTOKINES ,Adipose tissue ,Fibroblast growth factor ,0302 clinical medicine ,Adipocytes ,Immunology and Allergy ,Pharmacology (medical) ,Prostaglandin E2 ,Macrophage inflammatory protein ,Cells, Cultured ,CARTILAGE METABOLISM ,0303 health sciences ,HUMAN ARTICULAR CHONDROCYTES ,ROLES ,biology ,Synovial Membrane ,DIFFERENTIATION ,Female ,medicine.symptom ,Chemokines ,medicine.drug ,EXPRESSION ,medicine.medical_specialty ,Immunology ,CCL3 ,Down-Regulation ,Inflammation ,Dinoprostone ,03 medical and health sciences ,Chondrocytes ,Rheumatology ,Internal medicine ,Osteoarthritis ,medicine ,Humans ,030304 developmental biology ,Aged ,business.industry ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Coculture Techniques ,Endocrinology ,Gene Expression Regulation ,STROMAL CELLS ,TISSUE ,biology.protein ,INTERLEUKIN-1-BETA ,business ,MATRIX ,030217 neurology & neurosurgery ,Biomarkers - Abstract
Objective To examine the effect of different sources of Good Manufacturing Practice clinical grade adipose-derived mesenchymal stem cells (AD-MSCs) on inflammatory factors in osteoarthritic (OA) chondrocytes and synoviocytes. Methods AD-MSCs from infrapatellar Hoffa fat, subcutaneous (SC) hip fat, and SC abdominal fat were cocultured in Transwells with chondrocytes or synoviocytes. Inflammatory factors (interleukin-1β [IL-1β], tumor necrosis factor α, IL-6, CXCL1/growth-related oncogene α, CXCL8/IL-8, CCL2/monocyte chemotactic protein 1, CCL3/macrophage inflammatory protein 1α, and CCL5/RANTES) were evaluated by quantitative reverse transcription–polymerase chain reaction or multiplex bead–based immunoassay. The role of different immunomodulators was analyzed. Results All the inflammatory factors analyzed were down-modulated at the messenger RNA or protein level independently by all 3 AD-MSC sources or by allogeneic AD-MSCs used in coculture with chondrocytes or synoviocytes. Inflammatory factor down-modulation was observed only when AD-MSCs were cocultured with chondrocytes or synoviocytes that produced high levels of inflammatory factors, but no effect was observed in cells that produced low levels of those factors, thus highlighting a dependence of the AD-MSC effect on existing inflammation. The immunomodulators IL-10, IL-1 receptor antagonist, fibroblast growth factor 2, indoleamine 2,3-dioxygenase 1, and galectin 1 were not involved in AD-MSC effects, whereas the cyclooxygenase 2 (COX-2)/prostaglandin E2 (PGE2) pathway exerted a role in the mechanism of antiinflammatory AD-MSC action. Conclusion The antiinflammatory effects of AD-MSCs are probably not dependent on AD-MSC adipose tissue sources and donors but rather on the inflammatory status of OA chondrocytes and synoviocytes. AD-MSCs seem to be able to sense and respond to the local environment. Even though a combination of different molecules may be involved in AD-MSC effects, the COX-2/PGE2 pathway may play a role, suggesting that AD-MSCs may be useful for therapies in osteoarticular diseases.
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- 2013
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21. Intramyocardial transplantation of mesenchymal stromal cells for chronic myocardial ischemia and impaired left ventricular function: Results of the MESAMI 1 pilot trial
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Daniel Cussac, Gilles Lande, Nicolas Piriou, Joffrey Pozzo, Matthieu Berry, J.B. Ruidavet, Angelo Parini, Philippe Bourin, T. Le Tourneau, M. Lebrin, A. Huynh, S. Kramer, Olivier Lairez, Fabian Gross, Luc Sensebé, Alain Manrique, Damien Guijarro, Jean-Noël Trochu, Patricia Lemarchand, Jerome Roncalli, Michel Galinier, Guillaume Lamirault, Meyer Elbaz, unité de recherche de l'institut du thorax UMR1087 UMR6291 (ITX), Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS), Institut des Maladies Métaboliques et Cardiovasculaires (I2MC), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de thérapie cellulaire, Etablissement Français du Sang, Laboratoire de chimie organique et organométallique (LCOO), Université Sciences et Technologies - Bordeaux 1-Centre National de la Recherche Scientifique (CNRS), Equipe 7 Inserm U1048, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Toulouse III - Paul Sabatier (UT3), STROMALab, Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Etablissement Français du Sang-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Biothérapie, CHU Toulouse [Toulouse], CIC - Nantes, Université de Nantes (UN)-IFR26-Institut National de la Santé et de la Recherche Médicale (INSERM), Service de médecine nucléaire [CHU Caen], Université de Caen Normandie (UNICAEN), Normandie Université (NU)-Normandie Université (NU)-CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Tumorothèque de Caen Basse-Normandie (TCBN), Epidémiologie et analyses en santé publique : risques, maladies chroniques et handicaps (LEASP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées, Service de cardiologie [Toulouse], Hôpital de Rangueil, CHU Toulouse [Toulouse]-CHU Toulouse [Toulouse], Laboratoire national des champs magnétiques intenses - Grenoble (LNCMI-G ), Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Nantes - UFR de Médecine et des Techniques Médicales (UFR MEDECINE), Université de Nantes (UN)-Université de Nantes (UN), CHU Caen, Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Normandie Université (NU)-Tumorothèque de Caen Basse-Normandie (TCBN)-Université de Caen Normandie (UNICAEN), Normandie Université (NU), Institut du thorax, Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS), Semiconductor Photonics Research Group, Trinity College Dublin-Science Foundation Ireland-Enterprise Ireland-Higher Education Authority, Biosense Webster, Inc., Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-CHU Toulouse [Toulouse]-Hôpital de Rangueil, Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS), Lemarchand, Patricia, Unité de recherche de l'institut du thorax (ITX-lab), Université Sciences et Technologies - Bordeaux 1 (UB)-Centre National de la Recherche Scientifique (CNRS), Module Biothérapies, Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Service Cardiologie [CHU Toulouse], Pôle Cardiovasculaire et Métabolique [CHU Toulouse], Centre Hospitalier Universitaire de Toulouse (CHU Toulouse)-Centre Hospitalier Universitaire de Toulouse (CHU Toulouse), Institut National des Sciences Appliquées (INSA)-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées (INSA)-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Centre National de la Recherche Scientifique (CNRS)-Université Grenoble Alpes [2016-2019] (UGA [2016-2019]), Laboratoire national des champs magnétiques intenses - Grenoble (LNCMI), Centre National de la Recherche Scientifique (CNRS)-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National des Sciences Appliquées - Toulouse (INSA Toulouse), and Institut National des Sciences Appliquées (INSA)-Institut National des Sciences Appliquées (INSA)-Université Joseph Fourier - Grenoble 1 (UJF)-Université Grenoble Alpes (UGA)
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0301 basic medicine ,Cardiac function curve ,Male ,medicine.medical_specialty ,Single Photon Emission Computed Tomography Computed Tomography ,medicine.medical_treatment ,Myocardial Ischemia ,Pilot Projects ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,030204 cardiovascular system & hematology ,Revascularization ,Mesenchymal Stem Cell Transplantation ,Transplantation, Autologous ,03 medical and health sciences ,Ventricular Dysfunction, Left ,0302 clinical medicine ,Internal medicine ,Medicine ,Humans ,Prospective Studies ,[SDV.BC] Life Sciences [q-bio]/Cellular Biology ,ComputingMilieux_MISCELLANEOUS ,Cells, Cultured ,Ejection fraction ,Ischemic cardiomyopathy ,business.industry ,Myocardium ,Mesenchymal stem cell ,Middle Aged ,3. Good health ,Clinical trial ,Transplantation ,030104 developmental biology ,Treatment Outcome ,Cardiology ,Feasibility Studies ,Female ,Cardiology and Cardiovascular Medicine ,business ,Perfusion ,Follow-Up Studies - Abstract
Background: The MESAMI 1 trial was a bicentric pilot study designed to test the feasibility and safety of intramyocardially injected autologous bone marrow-derived mesenchymal stromal cells (MSCs) for the treatment of ischemic cardiomyopathy. Methods and Results: The study included 10 patients with chronic myocardial ischemia, left ventricular (LV) ejection fractions (EFs) of ≤35%, and reversible perfusion defects who were on stable optimal medical therapy and were not candidates for revascularization. MSCs (mean: 61.5×106 cells per patient) were injected into 10-16 viable sites at the border of the LV scar via a NOGA-guided catheter. Both primary endpoints, feasibility (successful harvest, expansion, and injection of autologous MSCs) and safety (absence of severe adverse events [SAEs]) were met in all 10 patients at the 1-month follow-up time point, and none of the SAEs reported during the full 2-year follow-up period were attributable to the study intervention. The results of secondary efficacy endpoint analyses identified significant improvements from baseline to Month 12 in LVEF (29.4±2.0% versus 35.7±2.5%; p=0.003), LV end-systolic volume (167.8±18.8 mL versus 156.1±28.6 mL; p=0.04), 6- minute walk test and NYHA functional class. Conclusions: Our results suggest that autologous MSCs can be safely administered to the hearts of patients with severe, chronic, reversible myocardial ischemia and impaired cardiac function and may be associated with improvements in cardiac performance, LV remodeling, and patient functional status. A randomized, double blind, multicenter, placebo-controlled clinical trial (MESAMI 2) will evaluate the efficacy of this treatment approach in a larger patient population. Clinical Trial Registration: Unique identifier: NCT01076920
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- 2016
22. Impact of rituximab on stem cell mobilization following ACVBP regimen in poor-risk patients with diffuse large B-cell lymphoma: results from a large cohort of patients
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Loic Isebaert, Christophe Fermé, Gilles Salles, Olivier Fitoussi, Suzanne Mathieu-Nafissi, Olivier Hequet, Philippe Bourin, Hervé Tilly, Alain Bohbot, Corinne Haioun, François Lefrère, Nicolas Mounier, Valérie Lapierre, Serge Bologna, Luc Fornecker, Dominique bastit-Barrau, and Jean-Michel Miclea
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Oncology ,medicine.medical_specialty ,Cyclophosphamide ,business.industry ,Immunology ,Hematology ,medicine.disease ,Lymphoma ,Surgery ,Regimen ,Autologous stem-cell transplantation ,Internal medicine ,medicine ,Immunology and Allergy ,Vindesine ,Rituximab ,business ,Diffuse large B-cell lymphoma ,Hematopoietic Stem Cell Mobilization ,medicine.drug - Abstract
BACKGROUND: The ACVBP regimen is an efficient induction regimen for poor-risk patients with diffuse large B-cell lymphoma (DLBCL) before consolidative autologous stem cell transplantation. Adjunction of the monoclonal anti-CD20 antibody rituximab (R-ACVBP) was recently found to be superior to ACVBP alone. This study assessed the impact of rituximab on stem cell mobilization in two similar consecutive groups of patients treated with ACVBP in two prospective, controlled trials. STUDY DESIGN AND METHODS: The first trial (LNH-98B-3) involved 137 patients treated with ACVBP alone. In the second trial (LNH-03-3B), 91 patients received an R-ACVBP regimen. Stem cell mobilization was performed after a course of (R)-ACVBP. RESULTS: The median peak numbers of blood CD34+ cell counts recorded before the first apheresis procedure in the ACVBP and R-ACVBP groups were 69 × 106 and 63 × 106/L, respectively (p = 0.55). The median numbers of CD34+ cells collected were 7.1 × 106 and 6.0 × 106 CD34+ cells/kg for the ACVBP and R-ACVBP groups, respectively (p = 0.13). The median number of apheresis procedures required for gathering the minimum amount of CD34+ cells (2 × 106/kg) was the same in the two groups. CONCLUSION: When compared with ACVBP alone, adjunction of rituximab does not impair stem cell mobilization.
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- 2012
23. Platelet lysate from whole blood-derived pooled platelet concentrates and apheresis-derived platelet concentrates for the isolation and expansion of human bone marrow mesenchymal stromal cells: production process, content and identification of active components
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Anita Ignatius, Caroline Maurer, Markus Rojewski, Mélanie Gadelorge, Ramin Lotfi, Sandrine Fleury-Cappellesso, Julia Dausend, Volker Mailänder, Daniel Fürst, Philippe Bourin, Natalie Fekete, Luc Sensebé, and Hubert Schrezenmeier
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Blood Platelets ,Cancer Research ,Stromal cell ,Immunology ,Becaplermin ,chemokines ,Bone Marrow Cells ,Biology ,03 medical and health sciences ,0302 clinical medicine ,Animals ,Humans ,Immunology and Allergy ,Platelet ,Genetics (clinical) ,mesenchymal progenitor cells ,Cell Proliferation ,030304 developmental biology ,Whole blood ,0303 health sciences ,Transplantation ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Proto-Oncogene Proteins c-sis ,Cell Biology ,Fibroblasts ,cytokines ,Recombinant Proteins ,3. Good health ,Apheresis ,Oncology ,030220 oncology & carcinogenesis ,Blood Component Removal ,Cancer research ,Original Article ,Fibroblast Growth Factor 2 ,Platelet lysate ,Stem cell ,mesenchymal stromal cells ,Fetal bovine serum - Abstract
Background aims The clinical use of human mesenchymal stromal cells (MSC) requires ex vivo expansion in media containing supplements such as fetal bovine serum or, alternatively, human platelet lysate (PL). Methods Platelet concentrates were frozen, quarantine stored, thawed and sterile filtered to obtain PL. PL content and its effect on fibroblast-colony-forming unit (CFU-F) formation, MSC proliferation and large-scale expansion were studied. Results PL contained high levels of basic fibroblast growth factor (bFGF), soluble CD40L (sCD40L), vascular cell adhesion molecule-1 (VCAM-1), intercellular adhesion molecule-1 (ICAM-1), platelet-derived growth factor AA (PDGF-AA), platelet-derived growth factor AB/BB (PDGF-AB/BB), chemokine (C-C) ligand 5 (CCL5; RANTES) transforming growth factor-β1 (TGF-β1) and chemokine (C-X-C) ligand 1/2/3 (GRO), with low batch-to-batch variability, and most were stable for up to 14 days. Inhibition of PDGF-BB and bFGF decreased MSC proliferation by about 20% and 50%, respectively. The strongest inhibition (about 75%) was observed with a combination of anti-bFGF + anti-PDGF-BB and anti-bFGF + anti-TGF-β1 + anti-PDGF-BB. Interestingly, various combinations of recombinant PDGF-BB, bFGF and TGF-β1 were not sufficient to promote cell proliferation. PL from whole blood-derived pooled platelet concentrates and apheresis platelet concentrates did not differ significantly in their growth-promoting activity on MSC. Conclusions PL enhances MSC proliferation and can be regarded as a safe tool for MSC expansion for clinical purposes. \in particular, PDGF-BB and bFGF are essential components for the growth-promoting effect of PL, but are not sufficient for MSC proliferation.
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- 2012
24. Bioactivity and Prognostic Significance of Growth Differentiation Factor GDF15 Secreted by Bone Marrow Mesenchymal Stem Cells in Multiple Myeloma
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Mélanie Gadelorge, Nicolas Espagnolle, Michel Attal, Philippe Moreau, Philippe Bourin, Hervé Avet-Loiseau, Jean-Jacques Fournié, Thierry Facon, Anne Huynh, Murielle Roussel, Pierre Cordelier, Jill Corre, Bernard Klein, Elodie Labat, and Benjamin Hebraud
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Adult ,Male ,Melphalan ,Cancer Research ,Growth Differentiation Factor 15 ,Antineoplastic Agents ,Bone Marrow Cells ,Bortezomib ,Cell Line, Tumor ,medicine ,Humans ,Neoplasm ,Lenalidomide ,Multiple myeloma ,Aged ,business.industry ,Growth differentiation factor ,Mesenchymal Stem Cells ,Middle Aged ,Prognosis ,medicine.disease ,Boronic Acids ,Thalidomide ,Oncology ,Drug Resistance, Neoplasm ,Cell culture ,Pyrazines ,Immunology ,Cancer research ,Female ,GDF15 ,Multiple Myeloma ,business ,Signal Transduction ,medicine.drug - Abstract
Overexpression of growth differentiation factor 15 (GDF15) by bone marrow mesenchymal stem cells occurs widely in patients with multiple myeloma, but the pathophysiologic effects of GDF15 in this setting remain undefined. GDF15 has been described in numerous solid tumors but never in hematologic malignancies. In this study, we report that GDF15 significantly increases survival of stroma-dependent multiple myeloma cells including primary multiple myeloma cells. In particular, GDF15 conferred resistance to melphalan, bortezomib, and to a lesser extent, lenalidomide in both stroma-dependent and stroma-independent multiple myeloma cells. Akt-dependent signaling was critical to mediate the effects of GDF15, whereas Src and extracellular signal-regulated kinase 1/2 signaling pathways were not involved. Given these results, we tested the clinical significance of plasma concentrations of GDF15 (pGDF15) in 131 patients with multiple myeloma and found that it correlated with disease prognosis. Specifically, patients with high levels of pGDF15 had lower probabilities of event-free and overall survival 30 months after diagnosis than patients with low pGDF15 levels. Our findings suggest that tumor microenvironment-derived GDF15 is a key survival and chemoprotective factor for multiple myeloma cells, which is pathophysiologically linked to both initial parameters of the disease as well as patient survival. Cancer Res; 72(6); 1395–406. ©2012 AACR.
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- 2012
25. Dual-energy X-ray absorptiometry and biochemical markers of bone turnover after autologous stem cell transplantation in myeloma
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Mélanie Gadelorge, Michel Attal, Muriel Roussel, Philippe Bourin, Jill Corre, Michel Laroche, and Olivia Lemaire
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Bone mineral ,Hyperparathyroidism ,medicine.medical_specialty ,medicine.diagnostic_test ,business.industry ,Hematology ,General Medicine ,medicine.disease ,Bone resorption ,Bone remodeling ,medicine.anatomical_structure ,Autologous stem-cell transplantation ,Endocrinology ,Internal medicine ,medicine ,business ,Dual-energy X-ray absorptiometry ,Multiple myeloma ,Femoral neck - Abstract
Objectives: To evaluate the effect of high-dose chemotherapy (HDT) followed by autologous stem cell transplantation (ASCT) on bone turnover and bone mineral density in a cohort of 39 consecutive patients with multiple myeloma (MM). Methods: Phosphorus and calcium parameters, bone turnover markers, and bone mineral density were studied. Timepoints were diagnosis (T1), just before ASCT (T2), 6 months (T3) after ASCT, and 1 yr (T4) after ASCT. Results: No bone mineral loss was shown on dual-energy X-ray absorptiometry (DXA) at T1 (lumbar Z-score −0.02, femoral neck Z-score 0.77) or during follow-up. Chronic vitamin D deficiency (25OHD3 11.7 ± 7.7 ng/mL at T1) and relative hyperparathyroidism from T2 to T4 were observed. In spite of this moderate hyperparathyroidism, serum C-telopeptide of type I collagen (CTX) decreased significantly between T1 and T4. Bone alkaline phosphatase levels were low at diagnosis and showed no significant change after ASCT, unlike DKK1 levels that were high at diagnosis and decreased 6 months after ASCT in patients not previously treated with bisphosphonates. Conclusion: Bone demineralization is moderate in multiple myeloma. ASCT induces a decrease in bone resorption but no changes in bone formation, remaining low despite the decrease in DKK1. Bone mineral loss, evaluated by DXA, is moderate in multiple myeloma. High-dose chemotherapy followed by ASCT leads to decreased bone resorption but osteoblastic bone formation remains low, in spite of reduced circulating DKK1.
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- 2012
26. Human Adipose-Derived Stromal Cells Efficiently Support Hematopoiesis In Vitro and In Vivo: A Key Step for Therapeutic Studies
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Philippe Bourin, Françoise Pflumio, Béatrice Cousin, Fabienne De Toni, Patrick Laharrague, Louis Casteilla, Sandrine Poglio, and Aissa Ben Youcef
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Adult ,Male ,Stromal cell ,Adipose tissue ,Antigens, CD34 ,Biology ,Mesenchymal Stem Cell Transplantation ,Subcutaneous fat ,Mice ,In vivo ,medicine ,Animals ,Humans ,Cell Lineage ,Aged ,Cell Proliferation ,Aged, 80 and over ,Mesenchymal stem cell ,Cell Differentiation ,Mesenchymal Stem Cells ,Cell Biology ,Hematology ,Middle Aged ,Hematopoietic Stem Cells ,In vitro ,Hematopoiesis ,Haematopoiesis ,medicine.anatomical_structure ,Adipose Tissue ,Immunology ,Cancer research ,Female ,Bone marrow ,Developmental Biology - Abstract
Adipose-derived stromal cells (ADSCs) are close relatives of bone marrow mesenchymal stromal cells (BM-MSCs). The ease of access to subcutaneous fat pad and the abundance of stromal precursors make fat tissue an attractive source of stromal cells for clinicians. However, their ability to support hematopoietic stem cells in vitro and in vivo has not been established definitively. Thus, their usefulness in supporting hematopoietic stem cell engraftment is not as clear as with BM-MSCs. In this article, we show that human ADSCs, cultured with a good manufacturing practice medium, maintain in vitro human early and committed hematopoietic progenitors and support their complete differentiation toward myeloid and lymphoid lineages. Compared with BM-MSCs, ADSCs elicit a more precocious early progenitor formation and faster proliferation and differentiation of hematopoietic progenitors. Further, in vivo, when co-injected in NOD.Cg-Prkdc(scid) Il2(rgtm1Wjl)/SzJ (NSG) mice with a low number of human CD34(+) cells, ADSCs enabled the higher production of immature human hematopoietic progenitors and CD45(+) cells when compared with BM-MSCs. As a whole, our results indicate that human ADSCs, isolated and expanded under clinical-grade conditions, support hematopoiesis in vitro and in vivo and thus provide the rationale for their use in supporting hematopoietic reconstitution in clinical settings.
- Published
- 2011
27. Cellules souches mésenchymateuses
- Author
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Philippe Bourin and Luc Sensebé
- Subjects
General Medicine ,General Biochemistry, Genetics and Molecular Biology - Abstract
Les cellules souches mesenchymateuses ou cellules stromales mesenchymateuses multipotentes (CSM) appartiennent a une population cellulaire initialement identifiee dans la moelle osseuse mais presente dans tous les tissus. Par leur potentiel de differenciation, leur production de cytokines, de facteurs trophiques et leurs actions immunosuppressives, les CSM sont un outil therapeutique tant en medecine regenerative que dans le traitement des pathologies immunitaires et inflammatoires. Actuellement, une centaine d’essais utilisant des CSM sont officiellement repertories. Un prerequis pour l’utilisation therapeutique des CSM est leur conformite avec les standards de « bonnes pratiques de fabrication » (BPF ou good manufacturing practices , GMP) assortis de controles de securite adequats. Le defi est de passer de procedes de culture utilises en recherche a des procedes de production correspondant a ces standards et a la reglementation nationale et internationale (europeenne et nord-americaine). Ceci necessite un travail de recherche et developpement en liaison directe avec les equipes de recherche et les equipes cliniques qui mettent en place les essais cliniques. C’est cette integration verticale, assurant des allers-retours permanents entre la recherche, le developpement et la clinique, qui permettra les developpements pertinents debouchant sur les procedes et les indications definitifs.
- Published
- 2011
28. Good Manufacturing Practices Production of Mesenchymal Stem/Stromal Cells
- Author
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Philippe Bourin, Luc Sensebé, Karin Tarte, Etablissement Français du Sang Centre-Atlantique, Etablissement Français du Sang Pyrénées-Méditerranée, Microenvironnement et cancer (MiCa), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), and Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique )
- Subjects
Quality Control ,Stromal cell ,MESH: Clinical Trials as Topic ,Mesenchyme ,MESH: Quality Control ,Cell Culture Techniques ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Bone Marrow Cells ,Biology ,Cell therapy ,MESH: Bioreactors ,03 medical and health sciences ,Bioreactors ,0302 clinical medicine ,Immune system ,Genetics ,medicine ,Humans ,Good manufacturing practice ,Molecular Biology ,MESH: Hematopoietic Stem Cell Transplantation ,030304 developmental biology ,MESH: Cell Culture Techniques ,Clinical Trials as Topic ,0303 health sciences ,MESH: Humans ,MESH: Bone Marrow Cells ,Mesenchymal stem cell ,Hematopoietic Stem Cell Transplantation ,Mesenchymal Stem Cells ,3. Good health ,Cell biology ,MESH: Mesenchymal Stem Cells ,medicine.anatomical_structure ,Cell culture ,030220 oncology & carcinogenesis ,Immunology ,Molecular Medicine ,MESH: Stromal Cells ,Stromal Cells ,Stem cell - Abstract
International audience; Because of their multi/pluripotency and immunosuppressive properties mesenchymal stem/stromal cells (MSCs) are important tools for treating immune disorders and for tissue repair. The increasing use of MSCs has led to production processes that need to be in accordance with Good Manufacturing Practice (GMP). In cellular therapy, safety remains one of the main concerns and refers to donor validation, choice of starting material, processes, and the controls used, not only at the batch release level but also during the development of processes. The culture processes should be reproducible, robust, and efficient. Moreover, they should be adapted to closed systems that are easy to use. Implementing controls during the manufacturing of clinical-grade MSCs is essential. The controls should ensure microbiological safety but also avoid potential side effects linked to genomic instability driving transformation and senescence or decrease of cell functions (immunoregulation, differentiation potential). In this rapidly evolving field, a new approach to controls is needed.
- Published
- 2011
29. Vascular and Endothelial Regeneration
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Patrick Laharrague, Valérie Planat-Benard, Louis Casteilla, Béatrice Cousin, and Philippe Bourin
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Pathology ,medicine.medical_specialty ,Stromal cell ,Induced Pluripotent Stem Cells ,Myocytes, Smooth Muscle ,Neovascularization, Physiologic ,Medicine (miscellaneous) ,Adipose tissue ,Biology ,Regenerative medicine ,chemistry.chemical_compound ,Vasculogenesis ,Ischemia ,Adipocyte ,medicine ,Animals ,Humans ,Induced pluripotent stem cell ,Mesenchymal stem cell ,General Medicine ,Cell biology ,medicine.anatomical_structure ,Adipose Tissue ,chemistry ,Cell Transdifferentiation ,Blood Vessels ,Bone marrow ,Stem Cell Transplantation - Abstract
Adipose tissue is the final tissue to develop and is strongly involved in energy homeostasis. It can represent up to 50% of body weight in obesity. Beside its metabolic role, endocrine functions appeared to play a key role in interconnecting adipose tissue with other tissues of the organism and in numerous physiological functions. The presence of adipocyte progenitors has long been demonstrated throughout life in the stromal fraction of adipose tissue. Now, it appears that these cells are multipotent and share numerous features with mesenchymal stem cells (MSC) derived from bone marrow. They also display some specificities and a strong pro-angiogenic potential. Altogether, these data emphasize the need to reconsider the potential of adipose tissue. Moreover, since fat pads are easy to sample, numerous and promising perspectives are now opening up in regenerative medicine, particularly in ischemic situations.
- Published
- 2010
30. Human Fibroblasts Share Immunosuppressive Properties with Bone Marrow Mesenchymal Stem Cells
- Author
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Philippe Bourin, Peter Winterton, Pol-André Apoil, Bénédicte Puissant-Lubrano, Sandrine Cappellesso-Fleury, Louis Casteilla, Matthias Titeux, and Antoine Blancher
- Subjects
Induced stem cells ,Stromal cell ,Chemistry ,Immunology ,Mesenchymal stem cell ,Immunology and Allergy ,Stem cell factor ,Lymphocyte proliferation ,Stem cell ,Stem cell transplantation for articular cartilage repair ,Cell biology ,GZMB - Abstract
Bone marrow mesenchymal stem cells (BM-MSCs) and adipose tissue-derived stem cells share immunosuppressive capacities, suggesting that the latter could be a general property of stromal cells. To check this hypothesis, we compared human BM-MSC and fibroblasts for their in vitro multi-potentiality, expandability and their immunomodulatory properties under normalized optimized culture conditions. We report that, unlike BM-MSCs, fibroblasts cannot differentiate in vitro into adipocytes and osteoblasts and differ from BM-MSCs by the expression of membrane CD106, CD10 and CD26 and by the expression of collagen VII mRNA. Like BM-MSCs, fibroblasts are unable to provoke in vitro allogeneic reactions, but strongly suppress lymphocyte proliferation induced by allogeneic mixed lymphocyte culture (MLC) or mitogens. We show that fibroblasts' immunosuppressive capacity is independent from prostaglandin E2, IL-10 and the tryptophan catabolising enzyme indoleamine 2,3-dioxygenase and is not abrogated after the depletion of CD8+ T lymphocytes, NK cells and monocytes. Finally, fibroblasts and BM-MSCs act at an early stage through blockage of lymphocyte activation, as demonstrated by down-regulation of GZMB (granzyme B) and IL2RA (CD25) expression.
- Published
- 2010
31. Mesenchymal stem cells for clinical application
- Author
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Mauro Krampera, Philippe Bourin, Luc Sensebé, Hubert Schrezenmeier, and Rosaria Giordano
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Adult ,Clinical uses of mesenchymal stem cells ,Biology ,Regenerative Medicine ,mesenchymal stem cells ,regenerative medicine ,immune regulation ,Regenerative medicine ,03 medical and health sciences ,0302 clinical medicine ,Humans ,030304 developmental biology ,Stem cell transplantation for articular cartilage repair ,0303 health sciences ,Multipotent Stem Cells ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Amniotic stem cells ,Hematology ,General Medicine ,3. Good health ,Cell biology ,Endothelial stem cell ,030220 oncology & carcinogenesis ,Immunology ,Stem cell ,Stem Cell Transplantation ,Adult stem cell - Abstract
Mesenchymal Stem Cells/Multipotent Marrow Stromal Cells (MSC) are multipotent adult stem cells present in all tissues, as part of the perivascular population. As multipotent cells, MSCs can differentiate into different tissues originating from mesoderm ranging from bone and cartilage, to cardiac muscle. Conflicting data show that MSCs could be pluripotent and able to differentiate into tissues and cells of non-mesodermic origin as neurons or epithelial cells. Moreover, MSCs exhibit non-HLA restricted immunosuppressive properties. This wide range of properties leads to increasing uses of MSC for immunomodulation or tissue repair. Based on their immunosuppressive properties MSC are used particularly in the treatment of graft versus host disease, For tissue repair, MSCs can work by different ways from cell replacement to paracrine effects through the release of cytokines and to regulation of immune/inflammatory responses. In regenerative medicine, trials are in progress or planed for healing/repair of different tissue or organs as bone, cartilage, vessels, myocardium, or epithelia. Although it has been demonstrated that ex-vivo expansion processes using fetal bovine serum, recombinant growth factors (e.g. FGF2) or platelet lysate are feasible, definitive standards to produce clinical-grade MSC are still lacking. MSCs have to be produced according GMP and regulation constraints. For answering to the numerous challenges in this fast developing field of biology and medicine, integrative networks linking together research teams, cell therapy laboratories and clinical teams are needed.
- Published
- 2010
32. Mesenchymal Stem Cells Promote Matrix Metalloproteinase Secretion by Cardiac Fibroblasts and Reduce Cardiac Ventricular Fibrosis After Myocardial Infarction
- Author
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Philippe Bourin, Denis Calise, Marie-Hélène Seguelas, Catherine Ordener, Angelo Parini, Olivier Lairez, Céline Mias, Jerome Roncalli, Daniel Cussac, Nathalie Augé, Elodie Trouche, Marie-Dominique Piercecchi-Marti, Anne Negre Salvayre, Institut de médecine moléculaire de Rangueil (I2MR), Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-IFR150-Institut National de la Santé et de la Recherche Médicale (INSERM), Zootechny, Department of Experimental Micro-Surgery, Physiopathologie de l'Endothelium, Vascular research center of Marseille (VRCM), Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de thérapie cellulaire (EFS), Etablissement Français du Sang, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées- Institut Fédératif de Recherche Bio-médicale Institution (IFR150)-Institut National de la Santé et de la Recherche Médicale (INSERM), Simon, Marie Francoise, Université de Toulouse (UT)-Université de Toulouse (UT)- Institut Fédératif de Recherche Bio-médicale Institution (IFR150)-Institut National de la Santé et de la Recherche Médicale (INSERM), and Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Aix Marseille Université (AMU)
- Subjects
MMP2 ,Cell Survival ,Cardiac fibrosis ,Heart Ventricles ,Blotting, Western ,Myocardial Infarction ,030204 cardiovascular system & hematology ,Matrix metalloproteinase ,Biology ,Mesenchymal Stem Cell Transplantation ,Polymerase Chain Reaction ,Extracellular matrix ,Mice ,03 medical and health sciences ,0302 clinical medicine ,In vivo ,Quantum Dots ,medicine ,Animals ,Collagenases ,Cells, Cultured ,030304 developmental biology ,Mice, Knockout ,Tissue Inhibitor of Metalloproteinase-2 ,0303 health sciences ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Cell Biology ,Anatomy ,Fibroblasts ,medicine.disease ,Fibrosis ,Immunohistochemistry ,Actins ,Rats ,Transplantation ,Matrix Metalloproteinase 9 ,Echocardiography ,Culture Media, Conditioned ,Heart failure ,Cancer research ,Matrix Metalloproteinase 2 ,Molecular Medicine ,Collagen ,Matrix Metalloproteinase 1 ,Developmental Biology - Abstract
Recent studies showed that mesenchymal stem cells (MSCs) transplantation significantly decreased cardiac fibrosis; however, the mechanisms involved in these effects are still poorly understood. In this work, we investigated whether the antifibrotic properties of MSCs involve the regulation of matrix metalloproteinases (MMPs) and matrix metalloproteinase endogenous inhibitor (TIMP) production by cardiac fibroblasts. In vitro experiments showed that conditioned medium from MSCs decreased viability, α-smooth muscle actin expression, and collagen secretion of cardiac fibroblasts. These effects were concomitant with the stimulation of MMP-2/MMP-9 activities and membrane type 1 MMP expression. Experiments performed with fibroblasts from MMP2-knockout mice demonstrated that MMP-2 plays a preponderant role in preventing collagen accumulation upon incubation with conditioned medium from MSCs. We found that MSC-conditioned medium also decreased the expression of TIMP2 in cardiac fibroblasts. In vivo studies showed that intracardiac injection of MSCs in a rat model of postischemic heart failure induced a significant decrease in ventricular fibrosis. This effect was associated with the improvement of morphological and functional cardiac parameters. In conclusion, we showed that MSCs modulate the phenotype of cardiac fibroblasts and their ability to degrade extracellular matrix. These properties of MSCs open new perspectives for understanding the mechanisms of action of MSCs and anticipate their potential therapeutic or side effects. Disclosure of potential conflicts of interest is found at the end of this article.
- Published
- 2009
33. Mesenchymal Stem Cells for Therapeutic Purposes
- Author
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Philippe Bourin and Luc Sensebé
- Subjects
Quality Control ,Transplantation ,business.industry ,Mesenchymal stem cell ,Cell Culture Techniques ,Adipose tissue ,Clinical uses of mesenchymal stem cells ,Mesenchymal Stem Cells ,Mesenchymal Stem Cell Transplantation ,Regenerative medicine ,Culture Media ,Cell biology ,Mesoderm ,medicine.anatomical_structure ,Antigens, CD ,Cord blood ,Immunology ,Cell Adhesion ,medicine ,Humans ,Bone marrow ,Stem cell ,business ,Cell Division ,Adult stem cell - Abstract
Mesenchymal stem cells (MSC) are multipotent adult stem cells harboring a wide range of differentiations and non-human leukocyte antigen-restricted immunosuppressive properties that lead to an increasing use of MSC in immunomodulation and in regenerative medicine. To produce MSC, definitive standards are still lacking. Whatever the starting material used (e.g., bone marrow, adipose tissue, or cord blood), numerous parameters including cell plating density, number of passages, and culture medium, play a major role in the culture process and have to be determined. To date, the different production processes have been effective, and based on phenotypic analysis and differentiation potential, a first set of simple controls have been defined. However, controls of the final product should provide precise data on efficacy and safety. The next challenge will be to develop production processes that reach good manufacturing practices goals and to define more accurate control methods of cultivated MSC.
- Published
- 2009
34. Low O2 concentrations enhance the positive effect of IL-17 on the maintenance of erythroid progenitors during co-culture of CD34+ and mesenchymal stem cells
- Author
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Pascale Duchez, Pavle Milenković, Jean Chevaleyre, Diana Bugarski, Philippe Bourin, Marija Vlaski, Zoran Ivanovic, Mohammad Hammoud, Jean-Michel Boiron, Vincent Praloran, Gordana Jovčić, and Aleksandra Krstić
- Subjects
Stromal cell ,Clinical Biochemistry ,Immunology ,CD34 ,Antigens, CD34 ,Stimulation ,Biology ,Colony-Forming Units Assay ,Humans ,Immunology and Allergy ,hematopoietic progenitors ,Erythroid Precursor Cells ,hypoxia ,Interleukin-6 ,Interleukin-17 ,Mesenchymal stem cell ,Mesenchymal Stem Cells ,Phenotype ,Cell Hypoxia ,Coculture Techniques ,Hematopoiesis ,Cell biology ,Oxygen ,IL-17 ,Haematopoiesis ,Limiting oxygen concentration ,Interleukin 17 ,oxygen ,CD34+ - Abstract
Co-culture of haematopoietic cells with a stromal cell layer does not mimic the physiological, micro-environmental niche, whose major feature is a low oxygen (O-2) concentration. Thus, in order to study the effects of IL-17 in a context which better approximates the physiological state, we investigated its effects on cell expansion, colony-forming ability, and the phenotypical profile of normal, human blood CD34(+) cells co-cultured for five days with MSC layers at various O-2 concentrations (20%, 12.5% and 3% O-2). We demonstrated that IL-17 enhances CD34(+) and total CFC production during the five days of MSC/CD34(+) co-culture. This effect depends upon the O-2 concentration, reaching its maximum at 3% O-2, and is more pronounced on erythroid progenitors (BFU-E). In addition, the stimulation of IL-6 production by IL-17 in MSC cultures and co-cultures is enhanced by low O-2 concentration. The expression of some differentiation markers (CD34, CD13 and CD41) on haematopoietic cells in co-cultures also depends upon the oxygen concentration. Our results strengthen the concept that physiological levels of O-2 (mistakenly called hypoxia), should be considered as an important environmental factor that significantly influences cytokine activity.
- Published
- 2009
35. A regulatory cross-talk between Vγ9Vδ2 T lymphocytes and mesenchymal stem cells
- Author
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Ludovic Martinet, Mélanie Gadelorge, Sandrine Fleury-Cappellesso, Jean-Jacques Fournié, Rémy Poupot, Gilles Dietrich, and Philippe Bourin
- Subjects
medicine.medical_treatment ,Immunology ,Mesenchymal stem cell ,Immunotherapy ,Biology ,Cell biology ,Cytokine ,medicine ,Immunology and Allergy ,Cytotoxic T cell ,Tumor necrosis factor alpha ,Interferon gamma ,IL-2 receptor ,Progenitor cell ,medicine.drug - Abstract
The physiological functions of human TCRVgamma9Vdelta2(+) gammadelta lymphocytes reactive to non-peptide phosphoantigens contribute to cancer immunosurveillance and immunotherapy. However, their regulation by mesenchymal stem cells (MSC), multipotent and immunomodulatory progenitor cells able to infiltrate tumors, has not been investigated so far. By analyzing freshly isolated TCRVgamma9Vdelta2(+) lymphocytes and primary cell lines stimulated with synthetic phosphoantigen or B-cell lymphoma cell lines in the presence of MSC, we demonstrated that MSC were potent suppressors of gammadelta-cell proliferation, cytokine production and cytolytic responses in vitro. This inhibition was mediated by the COX-2-dependent production of prostaglandin E2 (PGE(2)) and by MSC through EP2 and EP4 inhibitory receptors expressed by Vgamma9Vdelta2 T lymphocytes. COX-2 expression and PGE(2) production by MSC were not constitutive, but were induced by IFN-gamma and TNF-alpha secreted by activated Vgamma9Vdelta2 T cells. This regulatory cross-talk between MSC and Vgamma9Vdelta2 T lymphocytes involving PGE(2) could be of importance for the antitumor and antimicrobial activities of gammadelta T cells.
- Published
- 2009
36. Mesenchymal Progenitor Cells: Tissue Origin, Isolation And Culture
- Author
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Sandrine Fleury-Cappellesso, Mélanie Gadelorge, Christine Rage, Julie-Anne Peyrafitte, Marilyn Gomez, Philippe Bourin, and Luc Sensebé
- Subjects
Pathology ,medicine.medical_specialty ,business.industry ,Mesenchymal stem cell ,Review Article · Übersichtsarbeit ,Multipotent Mesenchymal Stromal Cells ,Hematology ,Isolation (microbiology) ,Cell biology ,Cell culture ,Immunology and Allergy ,Medicine ,Progenitor cell ,business - Abstract
SUMMARY: Since the pioneering work of Alexander Friedenstein on multipotent mesenchymal stromal cells (MSCs), a tremendous amount of work has been done to isolate, characterize and culture such cells. Assay of colony forming unit-fibroblasts (CFU-Fs), the hallmark of MSCs, is used to estimate their frequency in tissue. MSCs are adherent cells, so they are easy to isolate, and they show contact inhibition. Thus, several parameters must be taken into account for culture: cell density, number of passages, culture medium, and growth factors used. The purity of the initial material is not a limiting parameter. Similar but not identical cell populations are found in almost all mammal or human tissues. MSCs seem to be very abundant in adipose tissue but at low frequency in blood from umbilical cord or in adult tissue. The culture conditions are very similar, whatever the source of cells. Because of their favorable properties, MSCs are very promising tools for regenerative medicine.
- Published
- 2008
37. Producing MSC according GMP: Process and controls
- Author
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Philippe Bourin and Luc Sensebé
- Subjects
Tissue Engineering ,business.industry ,Mesenchymal stem cell ,Cell Culture Techniques ,Biomedical Engineering ,Mesenchymal Stem Cells ,General Medicine ,medicine.disease ,Biomaterials ,Transplantation ,Haematopoiesis ,Paracrine signalling ,Graft-versus-host disease ,Cancer research ,medicine ,Animals ,Humans ,Progenitor cell ,Stem cell ,business ,Adult stem cell - Abstract
The concept of progenitors/stem cells for cells of mesenchymal origin, which led to the description of mesenchymal stem cells (MSCs), arose during the 1970s with Friedenstein and colleagues [1]. Since the 1990s, increasing experimental data has allowed for the definition of MSCs as multipotent adult stem cells that can differentiate into connective skeletal tissue, bone, cartilage, marrow stroma and adipocytes [2–4]. In addition, controversial data suggest that MSCs may give rise to sarcomeric muscle (skeletal and cardiac) [5–7], endothelial cells [8], and even cells of nonmesodermal origin, such as hepatocytes [9], neural cells [10] and epithelial cells [11,12]. MSCs have immunoregulatory actions and interact with all the cells involved in immune response [13], different molecules or cytokines have been shown to play a role in this immuno-regulatory action [14, 15]. However, some controversial studies argue that MSCs could be immunogenic in immuno-competent mice suggesting that these cells are not intrinsically immuno-privileged [16]. The wide range of differentiation potential of MSCs, the possibility of their engraftment [17], their immunosuppressive effect [18], and their expansion through culture led to increasing clinical interest in the use of MSCs, through intravenous infusion or site-directed administration, in numerous pathologic situations. Since the first phase-1 study that demonstrated the good tolerance and safety of MSC transplantation [19], clinical trials have investigated large bone defects [20], genetic bone disease as osteogenesis imperfecta [17,21], haematopoietic stem cell transplantation for repair of haematopoietic stroma [22], treatment or prevention of graft versus host disease [23,24]. During the repair process of damaged tissues, the effects of MSCs can be related to different mechanisms, differentiation towards tissue specific pathway, repair of microenvironment with paracrine/juxtacrine effects by growth factors and cytokines [25] or extracellular matrix reorganization [26]. To perform clinical trials billions rather than millions of MSCs, solely or linked to biomaterials, are needed. Producing MSCs for that purpose necessitates adhering to good manufacturing practices (GMP) to insure the delivery of a “cell drug” that is safe, reproducible and efficient. However, standards for the
- Published
- 2008
38. Bone marrow mesenchymal stem cells are abnormal in multiple myeloma
- Author
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Bernard Klein, Clotaire Danho, Patrick Laharrague, Anne Huynh, Sandrine Fleury-Cappellesso, Michel Attal, Jill Corre, Philippe Bourin, Karène Mahtouk, Thierry Rème, Mélanie Gadelorge, Laboratoire de Thérapie Cellulaire, EFS, Laboratoire d'Hématologie [Purpan], Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-CHU Toulouse [Toulouse]-Hôpital Purpan [Toulouse], CHU Toulouse [Toulouse], Immunopathologie des maladies tumorales et autoimmunes, Université Montpellier 1 (UM1)-IFR76-Institut National de la Santé et de la Recherche Médicale (INSERM), Groupe d'Etude des Cellules Souches Mésenchymateuses (GECSOM), and GECSOM
- Subjects
Male ,Cancer Research ,Pathology ,Cellular differentiation ,MESH: Hematopoiesis ,MESH: Multiple Myeloma ,0302 clinical medicine ,Multiple myeloma ,MESH: Aged ,MESH: Cytokines ,0303 health sciences ,MESH: Middle Aged ,MESH: Bone Marrow Cells ,Cell Differentiation ,Hematology ,Middle Aged ,3. Good health ,Haematopoiesis ,medicine.anatomical_structure ,Oncology ,030220 oncology & carcinogenesis ,Cytokines ,Female ,Stem cell ,Multiple Myeloma ,MESH: Cell Differentiation ,Adult ,medicine.medical_specialty ,Growth Differentiation Factor 15 ,Stromal cell ,Bone Marrow Cells ,Biology ,Article ,MESH: Gene Expression Profiling ,03 medical and health sciences ,MESH: Cell Proliferation ,medicine ,Humans ,[SDV.BBM]Life Sciences [q-bio]/Biochemistry, Molecular Biology ,Aged ,Cell Proliferation ,030304 developmental biology ,MESH: Osteoblasts ,MESH: Humans ,Osteoblasts ,Gene Expression Profiling ,Mesenchymal stem cell ,MESH: Adult ,Mesenchymal Stem Cells ,medicine.disease ,MESH: Male ,Hematopoiesis ,MESH: Mesenchymal Stem Cells ,DKK1 ,Bone marrow ,MESH: Female - Abstract
International audience; Recent literature suggested that cells of the microenvironment of tumors could be abnormal as well. To address this hypothesis in multiple myeloma (MM), we studied bone marrow mesenchymal stem cells (BMMSCs), the only long-lived cells of the bone marrow microenvironment, by gene expression profiling and phenotypic and functional studies in three groups of individuals: patients with MM, patients with monoclonal gamopathy of undefined significance (MGUS) and healthy age-matched subjects. Gene expression profile independently classified the BMMSCs of these individuals in a normal and in an MM group. MGUS BMMSCs were interspersed between these two groups. Among the 145 distinct genes differentially expressed in MM and normal BMMSCs, 46% may account for a tumor-microenvironment cross-talk. Known soluble factors implicated in MM pathophysiologic features (i.e. IL (interleukin)-6, DKK1) were revealed and new ones were found which are involved in angiogenesis, osteogenic differentiation or tumor growth. In particular, GDF15 was found to induce dose-dependent growth of MOLP-6, a stromal cell-dependent myeloma cell line. Functionally, MM BMMSCs induced an overgrowth of MOLP-6, and their capacity to differentiate into an osteoblastic lineage was impaired. Thus, MM BMMSCs are abnormal and could create a very efficient niche to support the survival and proliferation of the myeloma cells.
- Published
- 2007
39. Adipose Mesenchymal Stromal Cell-Based Therapy for Severe Osteoarthritis of the Knee: A Phase I Dose-Escalation Trial
- Author
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Louis Casteilla, Gina Lisignoli, Christian Jorgensen, Joachim H. X. Schrauth, Frank Barry, Sophie Domergue, Yves-Marie Pers, Philippe Bourin, Daniel Haddad, C. Cyteval, François Canovas, Danièle Noël, Luc Sensebé, Christophe Delfour, Lars Rackwitz, Rosanna Ferreira, Ulrich Noeth, Oliver Pullig, Sandrine Fleury, Cellules Souches, Plasticité Cellulaire, Médecine Régénératrice et Immunothérapies (IRMB), Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Montpellier (UM), Hôpital Lapeyronie [Montpellier] (CHU), Julius-Maximilians-Universität Würzburg (JMU), Département de Rhumatologie[Montpellier], Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier)-Hôpital Lapeyronie, CHU Montpellier, Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), National University of Ireland [Galway] (NUI Galway), Établissement Français du Sang Alpes-Méditerranée (EFS Alpes-Méditerranée), STROMALab, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS), EFS, Istituto Ortopedico Rizzoli Bologna, University of Würzburg, Hôpital Gui de Chauliac [Montpellier], The research leading to these results has received funding from the European Union Seventh Framework Programme FP7/2007-2013 under grant agreement 241719. Work in the laboratory INSERM U844 was also supported by the Inserm Institute, the University of Montpellier, and the Agence Nationale pour la Recherche for support of the national infrastructure: 'ECELLFRANCE: Development of a national adult mesenchymal stem cell based therapy platform., ANR-11-INBS-0005,ECELLFRANCE,Développement d'une Plateforme Nationale pour la médecine régénératrice(2011), European Project: 241719,EC:FP7:HEALTH,FP7-HEALTH-2009-single-stage,ADIPOA(2010), Philips, Alexandre, Infrastructures - Développement d'une Plateforme Nationale pour la médecine régénératrice - - ECELLFRANCE2011 - ANR-11-INBS-0005 - INBS - VALID, ADIPOSE DERIVED STROMAL CELLS FOR OSTEOARTHRITIS - ADIPOA - - EC:FP7:HEALTH2010-01-01 - 2014-06-30 - 241719 - VALID, Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS), Hôpital Gui de Chauliac [CHU Montpellier], Julius-Maximilians-Universität Würzburg [Wurtzbourg, Allemagne] (JMU), Université de Montpellier (UM)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre Hospitalier Régional Universitaire [Montpellier] (CHRU Montpellier), Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Etablissement Français du Sang-Institut National de la Santé et de la Recherche Médicale (INSERM), and Publica
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0301 basic medicine ,Male ,[SDV]Life Sciences [q-bio] ,Phases of clinical research ,Arthritis ,Cell Count ,Osteoarthritis ,multicenter ,Severity of Illness Index ,Injections, Intra-Articular ,Cell therapy ,stem-cells ,0302 clinical medicine ,Musculoskeletal disorder ,Human Clinical Article ,cartilage ,ComputingMilieux_MISCELLANEOUS ,030222 orthopedics ,General Medicine ,Middle Aged ,Osteoarthritis, Knee ,3. Good health ,[SDV] Life Sciences [q-bio] ,medicine.anatomical_structure ,Treatment Outcome ,Adipose Tissue ,Regenerative medicine ,Adipose mesenchymal stromal cells ,Female ,therapeutic potential ,medicine.medical_specialty ,hyaluronic-acid ,Placebo ,Mesenchymal Stem Cell Transplantation ,MESH: Adipose Tissue / cytology ,Adipose Tissue / transplantation ,Aged ,Humans ,Mesenchymal Stem Cell Transplantation* / adverse effects ,Mesenchymal Stem Cell Transplantation* / methods ,Mesenchymal Stem Cells / cytology ,Osteoarthritis, Knee / pathology ,Osteoarthritis, Knee / therapy ,diseases ,03 medical and health sciences ,Internal medicine ,medicine ,Adverse effect ,model ,criteria ,business.industry ,Cartilage ,intraarticular injection ,tissue ,Mesenchymal Stem Cells ,Cell Biology ,medicine.disease ,Surgery ,Therapeutic potential ,030104 developmental biology ,Intra-articular injection ,Phase I clinical trial ,business ,Developmental Biology - Abstract
Osteoarthritis (OA) is the most widespread musculoskeletal disorder in adults. It leads to cartilage damage associated with subchondral bone changes and synovial inflammation, causing pain and disability. The present study aimed at evaluating the safety of a dose-escalation protocol of intra-articular injected adipose-derived stromal cells (ASCs) in patients with knee OA, as well as clinical efficacy as secondary endpoint. A bicentric, uncontrolled, open phase I clinical trial was conducted in France and Germany with regulatory agency approval for ASC expansion procedure in both countries. From April 2012 to December 2013, 18 consecutive patients with symptomatic and severe knee OA were treated with a single intra-articular injection of autologous ASCs. The study design consisted of three consecutive cohorts (six patients each) with dose escalation: low dose (2 × 106 cells), medium dose (10 × 106), and high dose (50 × 106). The primary outcome parameter was safety evaluated by recording adverse events throughout the trial, and secondary parameters were pain and function subscales of the Western Ontario and McMaster Universities Arthritis Index. After 6 months of follow-up, the procedure was found to be safe, and no serious adverse events were reported. Four patients experienced transient knee joint pain and swelling after local injection. Interestingly, patients treated with low-dose ASCs experienced significant improvements in pain levels and function compared with baseline. Our data suggest that the intra-articular injection of ASCs is a safe therapeutic alternative to treat severe knee OA patients. A placebo-controlled double-blind phase IIb study is being initiated to assess clinical and structural efficacy. Significance Although this phase I study included a limited number of patients without a placebo arm, it showed that local injection of autologous adipose-derived stem cells was safe and well tolerated in patients with knee osteoarthritis. This study also provides encouraging preliminary evidence of efficacy. Larger and controlled long-term studies are now mandatory to confirm whether this new strategy of cell therapy can improve pain and induce structural benefit in osteoarthritis.
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- 2015
40. Effects of a ceramic biomaterial on immune modulatory properties and differentiation potential of human mesenchymal stromal cells of different origin
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Philippe Bourin, Fabien Guilloton, Frédéric Deschaseaux, Mariano Di Trapani, Karin Tarte, Mauro Krampera, Massimo Dominici, Rosaria Giordano, Luc Sensebé, Hubert Schrezenmeier, Giulio Bassi, Cédric Ménard, Stem Cell Research Laboratory, Università degli studi di Verona = University of Verona (UNIVR), Etablissement Français du Sang, EFS, CHU Pontchaillou [Rennes], STROMALab, Université Toulouse III - Paul Sabatier (UT3), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS), Institute for transfusion medicine and Immunogenetic Clinics for Aplastic Anaemia, Ulm medical school, Laboratoire de thérapie cellulaire, Microenvironnement et cancer (MiCa), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), University of Verona (UNIVR), Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Etablissement Français du Sang-Institut National de la Santé et de la Recherche Médicale (INSERM), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Università degli Studi di Verona, Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), and Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS)
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Calcium Phosphates ,Ceramics ,medicine.medical_treatment ,[SDV]Life Sciences [q-bio] ,Biomedical Engineering ,Bioengineering ,Biochemistry ,Biomaterials ,Immunomodulation ,Immune system ,Materials Testing ,medicine ,Humans ,Immunologic Factors ,Cells, Cultured ,biology ,Tissue Scaffolds ,Mesenchymal Stromal Cells ,Chemistry ,Mesenchymal stem cell ,Cell Differentiation ,Mesenchymal Stem Cells ,Immunotherapy ,Original Articles ,Equipment Design ,Cell biology ,Equipment Failure Analysis ,medicine.anatomical_structure ,Durapatite ,Bone morphogenetic protein 4 ,Immunology ,Bone Substitutes ,Osteocalcin ,biology.protein ,Tumor necrosis factor alpha ,Bone marrow ,Stem cell - Abstract
The aim of this study was to assess the immune modulatory properties of human mesenchymal stromal cells obtained from bone marrow (BM-MSCs), fat (ASCs), and cord blood (CB-MSCs) in the presence of a hydroxyapatite and tricalcium-phosphate (HA/TCP) biomaterial as a scaffold for MSC delivery. In resting conditions, a short-term culture with HA/TCP did not modulate the anti-apoptotic and suppressive features of the various MSC types toward T, B, and NK cells; in addition, when primed with inflammatory cytokines, MSCs similarly increased their suppressive capacities in the presence or absence of HA/TCP. The long-term culture of BM-MSCs with HA/TCP induced an osteoblast-like phenotype with upregulation of OSTERIX and OSTEOCALCIN, similar to what was obtained with dexamethasone and, to a higher extent, with bone morphogenetic protein 4 (BMP-4) treatment. MSC-derived osteoblasts did not trigger immune cell activation, but were less efficient than undifferentiated MSCs in inhibiting stimulated T and NK cells. Interestingly, their suppressive machinery included not only the activation of indoleamine-2,3 dioxygenase (IDO), which plays a central role in T-cell inhibition, but also cyclooxygenase-2 (COX-2) that was not significantly involved in the immune modulatory effect of human undifferentiated MSCs. Since COX-2 is significantly involved in bone healing, its induction by HA/TCP could also contribute to the therapeutic activity of MSCs for bone tissue engineering.
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- 2015
41. Bone Marrow Mesenchymal Stem Cells Suppress Lymphocyte Proliferation In Vitro but Fail to Prevent Graft-versus-Host Disease in Mice
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Frédéric Charlotte, Aurélie Trenado, Béatrice Levacher, Jean-Jacques Lataillade, Françoise Norol, Philippe Bourin, José L. Cohen, Xavier Holy, Muriel Sudres, Jean-Paul Vernant, David Klatzmann, and Sylvie Grégoire
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Graft Rejection ,T cell ,Immunology ,Graft vs Host Disease ,Mice, Transgenic ,Cell Separation ,Lymphocyte proliferation ,Biology ,Mesenchymal Stem Cell Transplantation ,Immunophenotyping ,Mice ,In vivo ,medicine ,Animals ,Immunology and Allergy ,Lymphocytes ,Cells, Cultured ,Bone Marrow Transplantation ,Cell Proliferation ,Stem cell transplantation for articular cartilage repair ,Immunosuppression Therapy ,Mice, Inbred BALB C ,Mesenchymal stem cell ,medicine.disease ,Mice, Inbred C57BL ,Transplantation ,Disease Models, Animal ,medicine.anatomical_structure ,Graft-versus-host disease ,Radiation Chimera ,Female ,Bone marrow ,Lymphocyte Culture Test, Mixed ,Spleen - Abstract
Several reports have suggested that mesenchymal stem cells (MSCs) could exert a potent immunosuppressive effect in vitro, and thus may have a therapeutic potential for T cell-dependent pathologies. We aimed to establish whether MSCs could be used to control graft-vs-host disease (GVHD), a major cause of morbidity and mortality after allogeneic hemopoietic stem cell transplantation. From C57BL/6 and BALB/c mouse bone marrow cells, we purified and expanded MSCs characterized by the lack of expression of CD45 and CD11b molecules, their typical spindle-shaped morphology, together with their ability to differentiate into osteogenic, chondrogenic, and adipogenic cells. These MSCs suppressed alloantigen-induced T cell proliferation in vitro in a dose-dependent manner, independently of their MHC haplotype. However, when MSCs were added to a bone marrow transplant at a MSCs:T cells ratio that provided a strong inhibition of the allogeneic responses in vitro, they yielded no clinical benefit on the incidence or severity of GVHD. The absence of clinical effect was not due to MSC rejection because they still could be detected in grafted animals, but rather to an absence of suppressive effect on donor T cell division in vivo. Thus, in these murine models, experimental data do not support a significant immunosuppressive effect of MSCs in vivo for the treatment of GVHD.
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- 2006
42. Proteomic Study of Galectin-1 Expression in Human Mesenchymal Stem Cells
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A. Marie, C. Doucet, Philippe Bourin, T. Kadri, I. Ernou, Didier Lutomski, Jean-Jacques Lataillade, Michel Caron, and Raymonde Joubert-Caron
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Proteomics ,Time Factors ,Galectin 1 ,Blotting, Western ,Molecular Sequence Data ,Clinical uses of mesenchymal stem cells ,Bone Marrow Cells ,Biology ,Immunophenotyping ,Immune system ,Image Processing, Computer-Assisted ,Humans ,Electrophoresis, Gel, Two-Dimensional ,Amino Acid Sequence ,Cells, Cultured ,Cell Proliferation ,Stem cell transplantation for articular cartilage repair ,Cell Membrane ,Mesenchymal stem cell ,Antibodies, Monoclonal ,Cell Differentiation ,Mesenchymal Stem Cells ,Cell Biology ,Hematology ,Flow Cytometry ,Hematopoietic Stem Cells ,Extracellular Matrix ,Cell biology ,Haematopoiesis ,Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization ,Immunology ,Galectin-1 ,Stem cell ,Protein Binding ,Developmental Biology - Abstract
Bone marrow-derived mesenchymal stem cells (MSCs) are known to interact with hematopoietic stem cells (HSCs) and immune cells, and are of potential interest to be used as therapeutic agents for enhancing allogenic hematopoietic engraftment and preventing graft-versus-host disease (GVHD). Galectin 1 (Gal1) belongs to a family of structurally related molecules expressed in many vertebrate tissues that exert their functions both by binding to glycoconjugates, and by interaction with protein partners. In this work using a proteomic approach, we looked for the presence and the localization of Gal1 in short- and long-term culture of human (h) hMSC. We first determined, that Gal1 is one of the major proteins expressed in hMSC. We futher demonstrated that its expression is maintained when hMSC are expanded through a subculturing process up to five passages. Moreover, Gal1 is secreted and found at the cell surface of MSC, participating in extra cellular matrix (ECM)-cell interactions. Given the immunomodulatory properties of Gal1, its potential involvement in immunological functions of hMSC could be suggested.
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- 2005
43. Stromal cell–derived factor 1 regulates primitive hematopoiesis by suppressing apoptosis and by promoting G0/G1 transition in CD34+ cells: evidence for an autocrine/paracrine mechanism
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Denis Clay, Claude Jasmin, Catherine Dupuy, Philippe Bourin, Francis Herodin, Marie-Caroline Le Bousse-Kerdilès, and Jean-Jacques Lataillade
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Receptors, CXCR4 ,Stromal cell ,Immunology ,Antigens, CD34 ,Apoptosis ,Stem cell factor ,Biology ,Resting Phase, Cell Cycle ,Biochemistry ,Phosphatidylinositol 3-Kinases ,Paracrine signalling ,Paracrine Communication ,Humans ,Progenitor cell ,Autocrine signalling ,Interphase ,Cell growth ,G1 Phase ,Cell Biology ,Hematology ,Cell cycle ,Hematopoietic Stem Cells ,Chemokine CXCL12 ,Hematopoiesis ,Cell biology ,Autocrine Communication ,Stem cell ,Chemokines, CXC - Abstract
The stromal cell-derived factor 1 (SDF-1) chemokine has various effects on hematopoietic cell functions. Its role in migration and homing of hematopoietic progenitors is currently well established. Previously it was shown that SDF-1 stimulates myeloid progenitor proliferation in synergy with cytokines. Results of this study indicate that SDF-1 alone promotes survival of purified CD34(+) cells from human unmobilized peripheral blood (PB) by counteracting apoptosis as demonstrated by its capacity to reduce DNA fragmentation, annexin-V(+) cell number, and APO2.7 detection and to modulate bcl-2 homolog protein expression. The study demonstrates that SDF-1, produced by sorted CD34(+)CD38(+) cells and over-released in response to cell damage, exerts an antiapoptotic effect on CD34(+) cells through an autocrine/paracrine regulatory loop. SDF-1 participates in the autonomous survival of circulating CD34(+) cells and its effect required activation of the phosphotidyl inositol 3 kinase (PI3-K)/Akt axis. Cell sorting based on Hoechst/pyroninY fluorescences shows that SDF-1 production is restricted to cycling CD34(+) cells. SDF-1 triggers G(0) quiescent cells in G(1) phase and, in synergy with thrombopoietin or Steel factor, makes CD34(+) cells progress through S+G(2)/M phases of cell cycle. By assessing sorted CD34(+)CD38(-) and CD34(+)CD38(+) in semisolid culture, the study demonstrates that SDF-1 promotes survival of clonogenic progenitors. In conclusion, the results are the first to indicate a role for endogenous SDF-1 in primitive hematopoiesis regulation as a survival and cell cycle priming factor for circulating CD34(+) cells. The proposal is made that SDF-1 may contribute to hematopoiesis homeostasis by participating in the autonomous survival and cycling of progenitors under physiologic conditions and by protecting them from cell aggression in stress situations.
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- 2002
44. In-vitro circadian rhythm of murine bone marrow progenitor production
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Jacques Beau, Francis Lévi, Arnaud F. Ledain, Philippe Bourin, and Dominique Mille
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Male ,Physiology ,Ratón ,Circadian clock ,In Vitro Techniques ,Biology ,Colony-Forming Units Assay ,Mice ,In vivo ,Physiology (medical) ,medicine ,Animals ,Humans ,Circadian rhythm ,Progenitor cell ,Chronotherapy ,Mice, Inbred BALB C ,Granulocyte-Macrophage Colony-Stimulating Factor ,Hematopoietic Stem Cells ,Recombinant Proteins ,In vitro ,Circadian Rhythm ,Hematopoiesis ,Cell biology ,Haematopoiesis ,medicine.anatomical_structure ,Immunology ,Bone marrow - Abstract
Hematopoietic processes display 24h rhythms both in rodents and in human beings. We hypothesized these rhythms to be in part generated by a circadian oscillator within the bone marrow. The ability of murine bone marrow granulo-monocytic (GM) precursors to form colonies following colony-stimulating factor (rm GM-CSF) exposure was investigated in liquid culture samples obtained every 3 h for a span of up to 198 h. The CFU-GM count varied rhythmically over the first 4 d of culture, with a reproducible maximum in the early morning hours, similar to that observed in vivo. These experiments provide the first evidence that bone marrow progenitors sustain in vitro circadian rhythmicity, and they demonstrate the presence of a circadian time-keeping system within these cells. The results support the potential usefulness of bone marrow cultures for investigating chronopharmacologic effects of anticancer drugs and cytokines on this target system.
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- 2002
45. EFFECTS OF A NOVEL CERAMIC BIOMATERIAL ON IMMUNE MODULATORY PROPERTIES AND DIFFERENTIATION POTENTIAL OF MESENCHYMAL STROMAL CELLS
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M. Di Trapani, Philippe Bourin, Markus Rojewski, Pierre Layrolle, Massimo Dominici, Rosaria Giordano, Giulio Bassi, Martina Midolo, Cédric Ménard, Roberta Carusone, Serge Baroth, Lorenza Lazzari, Fabien Guilloton, Luciano Pacelli, Karin Tarte, Luc Sensebe, Isabelle Bezier, Cristiana Lavazza, Mauro Krampera, Hubert Schrezenmeier, Frederic Deschaseaux, Eliana Amati, Stem Cell Research Laboratory, University of Verona (UNIVR), Microenvironnement et cancer (MiCa), Université de Rennes 1 (UR1), Université de Rennes (UNIV-RENNES)-Université de Rennes (UNIV-RENNES)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Etablissement Français du Sang Centre-Atlantique (EA3855), EFS, STROMALab, Centre National de la Recherche Scientifique (CNRS)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Institut National Polytechnique (Toulouse) (Toulouse INP), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Etablissement Français du Sang-Institut National de la Santé et de la Recherche Médicale (INSERM), Institute for transfusion medicine and Immunogenetic Clinics for Aplastic Anaemia, Ulm medical school, Institut für Transfusionsmedizin, Universität Ulm - Ulm University [Ulm, Allemagne], Institut für klinische Transfusionsmedizin und Immungenetik, DRK Blutspendedienst Baden-Württemberg—Hessen, Laboratoire d'ingénierie osteo-articulaire et dentaire (LIOAD), Université de Nantes (UN)-IFR26-Institut National de la Santé et de la Recherche Médicale (INSERM), Laboratoire de Thérapie Cellulaire, Groupe d'Etude des Cellules Souches Mésenchymateuses (GECSOM), GECSOM, Università degli Studi di Verona, Université Toulouse III - Paul Sabatier (UT3), Université Fédérale Toulouse Midi-Pyrénées-Université Fédérale Toulouse Midi-Pyrénées-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université Fédérale Toulouse Midi-Pyrénées-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS), Università degli studi di Verona = University of Verona (UNIVR), Université de Rennes (UR)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Structure Fédérative de Recherche en Biologie et Santé de Rennes ( Biosit : Biologie - Santé - Innovation Technologique ), Université de Toulouse (UT)-Université de Toulouse (UT)-Ecole Nationale Vétérinaire de Toulouse (ENVT), Université de Toulouse (UT)-Université de Toulouse (UT)-Institut National Polytechnique (Toulouse) (Toulouse INP), and Université de Toulouse (UT)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Etablissement Français du Sang-Centre National de la Recherche Scientifique (CNRS)
- Subjects
Cancer Research ,medicine.medical_specialty ,Immunology ,Bone healing ,[SDV.BC]Life Sciences [q-bio]/Cellular Biology ,Research & Experimental Medicine ,Proinflammatory cytokine ,03 medical and health sciences ,0302 clinical medicine ,Immune system ,Internal medicine ,medicine ,Immunology and Allergy ,Genetics (clinical) ,030304 developmental biology ,0303 health sciences ,Transplantation ,Hematology ,biology ,Chemistry ,Mesenchymal stem cell ,Cell Biology ,Cell biology ,medicine.anatomical_structure ,Oncology ,Biotechnology & Applied Microbiology ,030220 oncology & carcinogenesis ,Cord blood ,Osteocalcin ,biology.protein ,Bone marrow - Abstract
International audience; The aim of this study was to assess the immune modulatory properties of human mesenchymal stromal cells obtained from bone marrow (BM-MSCs), fat (ASCs) and cord blood (CB-MSCs) in the presence of a novel hydroxyapatite and tricalcium-phosphate (HA/TCP) biomaterial as scaffold for MSC delivery. In resting conditions, a short-term culture with HA/TCP did not modulate the anti-apoptotic and suppressive features of the various MSC types towards T, B and NK cells; in addition, when primed with inflammatory cytokines, MSC maintained or not on HA/TCP similarlyincreased their suppressive capacities. The long-term culture of BM-MSCs with HA/TCP induced an osteoblast-like phenotype with up-regulation of OSTERIX and OSTEOCALCIN, similarly to what obtained with dexamethasone and, to a higher extent, BMP-4 treatment. MSC-derived osteoblasts did not trigger immune cell activation, but were less efficient than undifferentiated MSCs in inhibiting stimulated T and NK cells. Interestingly, their suppressive machinery included not only the activation of IDO, which plays a central role in T-cell inhibition, but also COX-2 that was not significantly involved in immune modulatory effect of human undifferentiated MSCs. COX-2 is significantly involved in bone healing, suggesting that its induction by HA/TCP could also contribute to the therapeutic activity of MSC for bone tissue engineering.
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- 2014
46. A clinical scalable cryopreservation method of adipose tissue for reconstructive surgery assessed by stromal vascular fraction and mice studies
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Nicolas Espagnolle, Julie Orio, Jean-Pierre Chavoin, Philippe Bourin, Jean-Louis Grolleau-Raoux, Mélanie Gadelorge, Ignacio Garrido, Louis Casteilla, Guillaume de Bonnecaze, Benoit Chaput, and Valérie Planat
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Adult ,Male ,Reconstructive surgery ,medicine.medical_specialty ,medicine.medical_treatment ,Adipose tissue ,Mice, Nude ,Cryopreservation ,Mice ,Cryoprotective Agents ,medicine ,Animals ,Humans ,Cells, Cultured ,business.industry ,Anatomy ,Equipment Design ,Stromal vascular fraction ,Plastic Surgery Procedures ,Surgery ,Plastic surgery ,Adipose Tissue ,Liposuction ,Models, Animal ,Heterografts ,Female ,Stromal Cells ,business - Abstract
Adipose tissue is widely used in plastic surgery. The main obstacle is that it can be used only immediately after liposuction, while reconstruction often requires several procedures to achieve optimal results. This study aimed to develop a cryopreservation protocol directly applicable to clinical situations, allowing repetitive procedures without multiple tissue harvests.The authors first tested scalable bags suitable for therapeutic uses. All subsequent experiments were performed in those bags. The authors evaluated in vitro, on the basis of cell viability, cell number, phenotype, and stromal cell proliferation, the efficacy of six cryopreservation media composed of an external cryoprotectant (human albumin or hydroxylethyl starch) with or without an internal cryoprotectant (dimethyl sulfoxide). Two storage temperatures (-196°C and -80°C) were tested in vitro and in vivo (subcutaneous graft in 30 nude mice) with the selected medium.The combination of 5% dimethyl sulfoxide and 95% hydroxylethyl yielded in vitro results that were good and the most consistent. With this cryoprotective solution, the authors observed no significant difference in vitro for a storage period of 7 days. When the storage was extended to 1 month, the cell viability was decreased by 10 percent for both storage temperatures. The in vivo experiments assessed the superiority of cryopreservation at -80°C with less graft resorption (60 percent and 70 percent, respectively, for -80°C and -196°C) and less fibrosis.The study's protocol with a chemically defined cryoprotective solution, specific scalable bags constrained in an aluminum holder, and a storage temperature of -80°C is promising for long-term adipose tissue cryopreservation.
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- 2014
47. Transportation conditions for prompt use of Ex Vivo expanded and freshly harvested clinical-grade bone marrow mesenchymal stromal/stem cells for bone regeneration
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Massimo Dominici, Elena Veronesi, Enrique Gómez-Barrena, Philippe Bourin, Pierre Layrolle, Alba Murgia, Valeria Rasini, Maria-Pau Ginebra, Fabio Catani, Giulia Grisendi, Markus Rojewski, Jorge S. Burns, Tiziana Montemurro, Anna Caselli, Rosaria Giordano, Maria Serena Piccinno, Carmen Panaitescu, Paolo Paolucci, Luc Sensebé, Hubert Schrezenmeier, Universitat Politècnica de Catalunya. Departament de Ciència dels Materials i Enginyeria Metal·lúrgica, Universitat Politècnica de Catalunya. BBT - Biomaterials, Biomecànica i Enginyeria de Teixits, Inserm, and UAM. Departamento de Cirugía
- Subjects
Biomineralization ,Calcium Phosphates ,Bone Regeneration ,Adhesion [Engineering controlled terms] ,Medicine (miscellaneous) ,Transportation ,Stem cells ,Cell Separation ,LS3_12 ,Mice ,0302 clinical medicine ,Mice, Inbred NOD ,Osteogenesis ,Ossos ,0303 health sciences ,Stem cell ,Tissue Scaffolds ,Chemistry ,Cell Differentiation ,Transmission control protocol ,3. Good health ,Cell biology ,Extracellular Matrix ,medicine.anatomical_structure ,Enginyeria de teixits ,Materials biomèdics ,030220 oncology & carcinogenesis ,clinical trials for bone regeneration ,cGMP ,MSC ,Stromal cell ,Biomedical materials ,Medicina ,Cell Survival ,Preservation, Biological ,Biomedical Engineering ,Bioengineering ,Bone Marrow Cells ,Buffers ,Choristoma ,Enginyeria dels materials [Àrees temàtiques de la UPC] ,Article ,NO ,Biomaterials ,03 medical and health sciences ,In vivo ,medicine ,Cell Adhesion ,Animals ,Humans ,Tissue engineering ,Viability assay ,Bone regeneration ,Bone ,Ships ,030304 developmental biology ,Cell Proliferation ,Bones ,Mesenchymal stem cell ,Three dimensional ,Biological materials ,Mesenchymal Stem Cells ,Durapatite ,Bone marrow ,Cytology ,Ex vivo ,Biomedical engineering - Abstract
This is a copy of an article published in Tissue Engineering: Part C and it is available online at http://online.liebertpub.com, Successful preliminary studies have encouraged a more translational phase for stem cell research. Nevertheless, advances in the culture of human bone marrow-derived mesenchymal stromal/stem cells (hBM-MSC) and osteoconductive qualities of combined biomaterials can be undermined if necessary cell transportation procedures prove unviable. We aimed at evaluating the effect of transportation conditions on cell function, including the ability to form bone in vivo, using procedures suited to clinical application. hBM-MSC expanded in current Good Manufacturing Practice (cGMP) facilities (cGMP-hBM-MSC) to numbers suitable for therapy were transported overnight within syringes and subsequently tested for viability. Scaled-down experiments mimicking shipment for 18 h at 4 C tested the influence of three different clinical-grade transportation buffers (0.9% saline alone or with 4% human serum albumin [HSA] from two independent sources) compared with cell maintenance medium. Cell viability after shipment was > 80% in all cases, enabling evaluation of (1) adhesion to plastic flasks and hydroxyapatite tricalcium phosphate osteoconductive biomaterial (HA/b-TCP 3D scaffold); (2) proliferation rate; (3) ex vivo osteogenic differentiation in contexts of 2D monolayers on plastic and 3D HA/b- TCP scaffolds; and (4) in vivo ectopic bone formation after subcutaneous implantation of cells with HA/b-TCP scaffold into NOD/SCID mice. Von Kossa staining was used to assess ex vivo osteogenic differentiation in 3D cultures, providing a quantifiable test of 3D biomineralization ex vivo as a rapid, cost-effective potency assay. Near-equivalent capacities for cell survival, proliferation, and osteogenic differentiation were found for all transportation buffers. Moreover, cGMP-hBM-MSC transported from a production facility under clinical-grade conditions of 4% HSA in 0.9% saline to a destination 18 h away showed prompt adhesion to HA/b-TCP 3D scaffold and subsequent in vivo bone formation. A successfully validated transportation protocol extends the applicability of fresh stem cells involving multicentric trials for regenerative medicine, This work was supported in part by the European Commission Seventh Framework Program (FP7/2007–2013) (grant no. 241879), through the REBORNE project, the Italian Ministry of Health ‘‘Bando Cellule Staminali 2008’’ (M.D., P.P.), Regione Emilia Romagna (P.P., M.D.), and the Associazione ASEOP (P.P.).
- Published
- 2014
48. Concise review: growth differentiation factor 15 in pathology: a clinical role?
- Author
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Philippe Bourin, Jill Corre, and Benjamin Hebraud
- Subjects
Pathology ,medicine.medical_specialty ,Erythrocytes ,Growth Differentiation Factor 15 ,Disease ,Biology ,Placenta ,Neoplasms ,medicine ,Biomarkers, Tumor ,Animals ,Humans ,Erythropoiesis ,Receptor ,Pathological ,Mechanism (biology) ,Cell Biology ,General Medicine ,Prognosis ,Tissue-Specific Progenitor and Stem Cells ,medicine.anatomical_structure ,Gene Expression Regulation ,Cardiovascular Diseases ,GDF15 ,Signal transduction ,Developmental Biology ,Transforming growth factor ,Signal Transduction - Abstract
Growth differentiation factor 15 (GDF15) is a divergent member of the transforming growth factor β family discovered in a broad range of cells, as indicated by the diversity of its nomenclature. However, the only tissue that expresses a high amount of GDF15 in the physiologic state is placenta. GDF15 is easily detected in blood, and its concentration varies with age. In fact, increased blood concentration of GDF15 is associated with numerous pathological conditions. However, the biological significance underlying these observations is far from clear. GDF15 could have a positive or negative role depending on the state of cells or their environment. Furthermore, study of its biology is hampered by lack of knowledge of its receptor and thus the signaling pathways that drive its action. GDF15 seems to be an integrative signal in pathologic conditions, giving information on severity of disease. Its effectiveness in classifying patients to modulate treatment remains to be shown. Development of therapeutic interventions with GDF15 or anti-GDF15 agents remains difficult until we uncover the mechanism that drives its activity.
- Published
- 2013
49. Regulation of CD45-induced signaling by galectin-1 in Burkitt lymphoma B cells
- Author
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Martine Raphael, Magali Fouillit, Michel Caron, Matthieu Levi-Strauss, Philippe Bourin, Dominique Bladier, Éva Monostori, Florence Poirier, and Raymonde Joubert-Caron
- Subjects
Galectin 1 ,Phosphatase ,Cell ,Protein tyrosine phosphatase ,Biochemistry ,Cell membrane ,LYN ,hemic and lymphatic diseases ,Phosphoprotein Phosphatases ,Tumor Cells, Cultured ,medicine ,Humans ,Phosphorylation ,Galectin ,B-Lymphocytes ,Chemistry ,Cell Membrane ,Flow Cytometry ,Burkitt Lymphoma ,Recombinant Proteins ,Cell biology ,carbohydrates (lipids) ,Hemagglutinins ,src-Family Kinases ,medicine.anatomical_structure ,Cell culture ,Leukocyte Common Antigens ,Signal transduction ,Signal Transduction - Abstract
It has been well established that Galectin-1 (GAL1), a beta-galactoside-binding protein, regulates the viability of lymphoid cells. However, the signaling pathway governed by the binding of GAL1 to the cell membrane is not understood. As a first step towards the elucidation of GAL1-initiated signaling events leading to a reduced viability of Burkitt lymphoma B cells, we tried to characterize the initial events induced by the binding of GAL1 to its receptor. This characterization was performed in BL36 cells, a Burkitt lymphoma cell line sensitive to GAL1. The results were as follows: (1) when solubilized cell membrane lysates were affinity bound to immobilized GAL1 and eluted by competition, the tyrosine phosphatase glyco-protein CD45 was found in the eluate, highlighting the role of CD45 as a receptor of GAL1; (2) the phosphatase activity of cell membranes diminished after incubation with GAL1; (3) immunoprecipitation experiments demonstrated that the phosphotyrosine kinase Lyn was dysregulated in cells that have been cultured in medium containing 700 nM GAL1, and (4) that the ratio between two isoforms of Lyn was modified during the treatment with GAL1. The regulation of Lyn therefore seems to be a key event in the action of GAL1.
- Published
- 2000
50. Chemokine SDF-1 enhances circulating CD34+ cell proliferation in synergy with cytokines: possible role in progenitor survival
- Author
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Claude Jasmin, Jean-Jacques Lataillade, Sylvain Rigal, Denis Clay, Marie-Caroline Le Bousse-Kerdilès, Catherine Dupuy, and Philippe Bourin
- Subjects
Adult ,Receptors, CXCR4 ,Chemokine ,Stromal cell ,Cell Survival ,Immunology ,Antigens, CD34 ,Hematopoietic Cell Growth Factors ,Biochemistry ,Colony-Forming Units Assay ,Cell therapy ,Transforming Growth Factor beta ,Humans ,Stromal cell-derived factor 1 ,Phosphorylation ,Progenitor cell ,Erythroid Precursor Cells ,Stem Cell Factor ,biology ,Cell growth ,Cell Cycle ,Antibodies, Monoclonal ,Granulocyte-Macrophage Colony-Stimulating Factor ,Drug Synergism ,Cell Biology ,Hematology ,Protein-Tyrosine Kinases ,Hematopoietic Stem Cells ,Chemokine CXCL12 ,Cell biology ,Haematopoiesis ,biology.protein ,Interleukin-3 ,Stem cell ,Chemokines, CXC ,Megakaryocytes ,Protein Processing, Post-Translational ,Cell Division - Abstract
The chemokine stromal cell-derived factor-1 (SDF-1), and its receptor, CXCR-4, have been implicated in the homing and mobilization of human CD34(+) cells. We show here that SDF-1 may also be involved in hematopoiesis, promoting the proliferation of human CD34(+) cells purified from normal adult peripheral blood (PB). CXCR-4 was expressed on PB CD34(+) cells. The amount of CXCR-4 on PB CD34(+) cells was 10 times higher when CD34(+) cells were purified following overnight incubation. CXCR-4 overexpression was correlated with a primitive PB CD34(+) cell subset defined by a CD34(high) CD38(low)CD71(low)c-Kit(low)Thy-1(+) antigenic profile. The functional significance of CXCR-4 expression was ascertained by assessing the promoting effect of SDF-1alpha on cell cycle, proliferation, and colony formation. SDF-1 alone increased the percentage of CD34(+) cells in the S+G(2)/M phases and sustained their survival. In synergy with cytokines, SDF-1 increased PB CD34(+) and CD34(high)CD38(low) cell expansion and colony formation. SDF-1 also stimulated the growth of colonies derived from primitive progenitors released from quiescence by anti-TGF-beta treatment. Thus, our results shed new light on the potential role of this chemokine in the stem cell engraftment process, which involves migration, adhesion, and proliferation. Furthermore, both adhesion-induced CXCR-4 overexpression and SDF-1 stimulating activity may be of clinical relevance for improving cell therapy settings in stem cell transplantation.
- Published
- 2000
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