18 results on '"Baksh D"'
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2. Mesenchymal Cell Culture
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Davies, John E., primary, Karp, J.M., additional, and Baksh, D., additional
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- 2002
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3. Mesenchymal stem cell-based cartilage tissue engineering: cells, scaffold and biology
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Song, L., Baksh, D., and Tuan, R.S.
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- 2004
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4. Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy
- Author
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Baksh, D., primary, Song, L., additional, and Tuan, R. S., additional
- Published
- 2004
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5. Manipulation of adult bone marrow derived stem and progenitor cell population. Dynamics in suspension culture
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Madlambayan, G.J., primary, Baksh, D., additional, Rogers, I., additional, Casper, R.F., additional, Davies, J.E., additional, and Zandstra, P.W., additional
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6. Manipulation of adult bone marrow derived stem and progenitor cell population. Dynamics in suspension culture.
- Author
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Madlambayan, G.J., Baksh, D., Rogers, I., Casper, R.F., Davies, J.E., and Zandstra, P.W.
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- 2002
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7. Chapter 26 - Mesenchymal Cell Culture: Bone
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Davies, John E., Karp, J.M., and Baksh, D.
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8. The importance of both fibroblasts and keratinocytes in a bilayered living cellular construct used in wound healing.
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Wojtowicz AM, Oliveira S, Carlson MW, Zawadzka A, Rousseau CF, and Baksh D
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- Basement Membrane, Cells, Cultured, Culture Media, Female, Humans, Male, Skin, Artificial, Structure-Activity Relationship, Varicose Ulcer immunology, Epidermal Cells, Fibroblasts metabolism, Keratinocytes metabolism, Varicose Ulcer pathology, Wound Healing
- Abstract
Cross talk between fibroblasts and keratinocytes, which maintains skin homeostasis, is disrupted in chronic wounds. For venous leg ulcers and diabetic foot ulcers, a bilayered living cellular construct (BLCC), containing both fibroblasts and keratinocytes that participate in cross talk, is a safe and effective product in healing chronic wounds. To show the importance of both cell types in BLCC, constructs were generated containing only fibroblasts or only keratinocytes and compared directly to BLCC via histology, mechanical testing, gene/protein analysis, and angiogenesis assays. BLCC contained a fully differentiated epithelium and showed greater tensile strength compared with one-cell-type constructs, most likely due to formation of intact basement membrane and well-established stratum corneum in BLCC. Furthermore, expression of important wound healing genes, cytokines, and growth factors was modulated by the cells in BLCC compared with constructs containing only one cell type. Finally, conditioned medium from BLCC promoted greater endothelial network formation compared with media from one-cell-type constructs. Overall, this study characterized a commercially available wound healing product and showed that the presence of both fibroblasts and keratinocytes in BLCC contributed to epithelial stratification, greater tensile strength, modulation of cytokine and growth factor expression, and increased angiogenic properties compared with constructs containing fibroblasts or keratinocytes alone., (© 2014 The Authors. Wound Repair and Regeneration published by Wiley Periodicals, Inc. on behalf of Wound Healing Society.)
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- 2014
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9. Culture of mesenchymal stem/progenitor cells in adhesion-independent conditions.
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Baksh D and Davies JE
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- Animals, Bioreactors, Cell Lineage, Cells, Cultured, Humans, Mesenchymal Stem Cells physiology, Phenotype, Stem Cells physiology, Cell Adhesion, Cell Culture Techniques instrumentation, Cell Culture Techniques methods, Mesenchymal Stem Cells cytology, Stem Cells cytology
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- 2008
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10. Human palatine tonsil: a new potential tissue source of multipotent mesenchymal progenitor cells.
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Janjanin S, Djouad F, Shanti RM, Baksh D, Gollapudi K, Prgomet D, Rackwitz L, Joshi AS, and Tuan RS
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- Adolescent, Adult, Biopsy, Bone Marrow Cells drug effects, Bone Marrow Cells metabolism, Cell Proliferation, Cell Survival physiology, Cells, Cultured, Child, Child, Preschool, Humans, Immunosuppression Therapy, Interferon-gamma pharmacology, Lymphocyte Culture Test, Mixed, Mesenchymal Stem Cells drug effects, Mesenchymal Stem Cells metabolism, Middle Aged, Multipotent Stem Cells drug effects, Multipotent Stem Cells metabolism, Palatine Tonsil drug effects, Palatine Tonsil metabolism, Receptors, Interferon metabolism, Interferon gamma Receptor, Bone Marrow Cells cytology, Mesenchymal Stem Cells cytology, Multipotent Stem Cells cytology, Palatine Tonsil cytology
- Abstract
Introduction: Mesenchymal progenitor cells (MPCs) are multipotent progenitor cells in adult tissues, for example, bone marrow (BM). Current challenges of clinical application of BM-derived MPCs include donor site morbidity and pain as well as low cell yields associated with an age-related decrease in cell number and differentiation potential, underscoring the need to identify alternative sources of MPCs. Recently, MPC sources have diversified; examples include adipose, placenta, umbilicus, trabecular bone, cartilage, and synovial tissue. In the present work, we report the presence of MPCs in human tonsillar tissue., Methods: We performed comparative and quantitative analyses of BM-MPCs with a subpopulation of adherent cells isolated from this lymphoid tissue, termed tonsil-derived MPCs (T-MPCs). The expression of surface markers was assessed by fluorescent-activated cell sorting analysis. Differentiation potential of T-MPCs was analyzed histochemically and by reverse transcription-polymerase chain reaction for the expression of lineage-related marker genes. The immunosuppressive properties of MPCs were determined in vitro in mixed lymphocyte reactions., Results: Surface epitope analysis revealed that T-MPCs were negative for CD14, CD31, CD34, and CD45 expression and positive for CD29, CD44, CD90, and CD105 expression, a characteristic phenotype of BM-MPCs. Similar to BM-MPCs, T-MPCs could be induced to undergo adipogenic differentiation and, to a lesser extent, osteogenic and chondrogenic differentiation. T-MPCs did not express class II major histocompatibility (MHC) antigens, and in a similar but less pronounced manner compared with BM-MPCs, T-MPCs were immunosuppressive, inhibiting the proliferation of T cells stimulated by allogeneic T cells or by non-specific mitogenic stimuli via an indoleamine 2,3-dioxygenase-dependent mechanism., Conclusion: Human palatine T-MPCs represent a new source of progenitor cells, potentially applicable for cell-based therapies.
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- 2008
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11. Isolation, characterization, and differentiation of human umbilical cord perivascular cells (HUCPVCs).
- Author
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Ennis J, Sarugaser R, Gomez A, Baksh D, and Davies JE
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- Cell Culture Techniques, Cells, Cultured, Humans, Blood Vessels cytology, Cell Differentiation, Cell Separation, Phenotype, Umbilical Cord cytology
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- 2008
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12. A non-contact suspension culture approach to the culture of osteogenic cells derived from a CD49elow subpopulation of human bone marrow-derived cells.
- Author
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Baksh D, Zandstra PW, and Davies JE
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- Cell Culture Techniques methods, Cells, Cultured, Flow Cytometry, Humans, Mesenchymal Stem Cells metabolism, Osteoblasts cytology, Bone Marrow Cells cytology, Cell Differentiation physiology, Integrin alpha5 metabolism, Mesenchymal Stem Cells cytology, Osteoblasts metabolism
- Abstract
We demonstrate that adult human bone marrow (BM) contains a population of mesenchymal stromal cells (MSCs) that can be expanded in non-adherent, cytokine-dependent, suspension culture conditions for at least 42 days. The cells generated during suspension culture lacked detectable levels of gene expression associated with differentiated mesenchymal cell types, including bone, muscle and fat, suggesting that suspension culture maintains MSCs in an uncommitted state. However, when these undifferentiated cells were taken out of suspension culture and placed in adherent osteogenic conditions, osteogenic genes were upregulated and morphologically identifiable bone matrix was elaborated. Flow cytometric analysis of uncultured, density gradient-separated human BM revealed that colony forming unit-fibroblast (CFU-F) and CFU-osteoblast (CFU-O) activity was associated with a CD45(-) CD49e(low) phenotype. Importantly, suspension-grown MSCs, capable of CFU-F and CFU-O development, maintained the CD45(-)CD49e(low) phenotype whereas MSCs directly cultured under adherent conditions rapidly upregulated CD49e expression and were associated with a CD45(-)CD49e(high) phenotype. Tracking the CD49e(low) expression under suspension culture conditions provides a mechanism to isolate an expanding suspension-grown MSC population with osteogenic potential. This could provide a potential strategy to isolate populations of MSCs, with functional osteogenic capacity, in a scalable and controllable culture system for therapeutic applications., (Copyright 2007 Wiley Periodicals, Inc.)
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- 2007
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13. Canonical and non-canonical Wnts differentially affect the development potential of primary isolate of human bone marrow mesenchymal stem cells.
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Baksh D and Tuan RS
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- Adult Stem Cells immunology, Aged, Bone Marrow Cells immunology, Cell Proliferation, Cells, Cultured, Colony-Forming Units Assay, Culture Media, Conditioned metabolism, Fibroblasts metabolism, Humans, Immunohistochemistry, Leukocyte Common Antigens analysis, Mesenchymal Stem Cells immunology, Microscopy, Fluorescence, Middle Aged, Osteoblasts metabolism, Proto-Oncogene Proteins genetics, Time Factors, Transfection, Wnt Proteins genetics, Wnt-5a Protein, Wnt3 Protein, Wnt3A Protein, Adult Stem Cells metabolism, Autocrine Communication, Bone Marrow Cells metabolism, Cell Differentiation, Mesenchymal Stem Cells metabolism, Paracrine Communication, Proto-Oncogene Proteins metabolism, Wnt Proteins metabolism
- Abstract
This study examines the role of Wnt signaling events in regulating the differential potential of mesenchymal stem cells (MSCs) from adult bone marrow (BM). Immunohistochemical analysis of BM revealed co-localization of Wnt5a protein, a non-canonical Wnt, with CD45(+) cells and CD45(-) STRO-1(+) cells, while Wnt3a expression, a canonical Wnt, was associated with the underlying stroma matrix, suggesting that Wnts may regulate MSCs in their niche in BM. To elucidate the role of Wnts in MSC development, adult human BM-derived mononuclear cells were maintained as suspension cultures to recapitulate the marrow cellular environment, in serum-free, with the addition of Wnt3a and Wnt5a protein. Results showed that Wnt3a increased cell numbers and expanded the pool of MSCs capable of colony forming unit -- fibroblast (CFU-F) and CFU -- osteoblast (O), while Wnt5a maintained cell numbers and CFU-F and CFU-O numbers. However, when cells were cultured directly onto tissue culture plastic, Wnt5a increased the number of CFU-O relative to control conditions. These findings suggest the potential dual role of Wnt5a in the maintenance of MSCs in BM and enhancing osteogenesis ex vivo. Our work provides evidence that Wnts can function as mesenchymal regulatory factors by providing instructive cues for the recruitment, maintenance, and differentiation of MSCs.
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- 2007
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14. Cross-talk between Wnt signaling pathways in human mesenchymal stem cells leads to functional antagonism during osteogenic differentiation.
- Author
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Baksh D, Boland GM, and Tuan RS
- Subjects
- Aged, Cell Proliferation, Culture Media, Conditioned, Frizzled Receptors metabolism, Genes, Reporter, Hepatocyte Nuclear Factor 1-alpha metabolism, Humans, LDL-Receptor Related Proteins metabolism, Low Density Lipoprotein Receptor-Related Protein-5, Middle Aged, Models, Biological, Osteocalcin genetics, Transcription, Genetic, Wnt-5a Protein, Wnt3 Protein, Wnt3A Protein, Cell Differentiation, Mesenchymal Stem Cells cytology, Mesenchymal Stem Cells metabolism, Osteogenesis, Proto-Oncogene Proteins metabolism, Signal Transduction, Wnt Proteins metabolism
- Abstract
Wnt signaling is involved in developmental processes and in adult stem cell homeostasis. This study analyzes the role(s) of key Wnt signaling mediators in the maintenance and osteogenesis of mesenchymal stem cells (MSCs). We focus specifically on the involvement of low-density lipoprotein-related protein 5 (LRP5), T-cell factor 1 (TCF1), and Frizzled (Fz) receptors, in the presence or absence of exogenous, prototypical canonical (Wnt3a), and non-canonical (Wnt5a) Wnts. In undifferentiated MSCs, LRP5 and TCF1 mediate canonical Wnt signal transduction, leading to increased proliferation, enhanced synergistically by Wnt3a. However, LRP5 overexpression inhibits osteogenic differentiation, further suppressed by Wnt3a. Wnt5a does not affect cell proliferation but enhances osteogenesis of MSCs. Interestingly, Wnt5a inhibits Wnt3a effects on MSCs, while Wnt3a suppresses Wnt5a-mediated enhancement of osteogenesis. Flow cytometry revealed that LRP5 expression elicits differential changes in Fz receptor profiles in undifferentiated versus osteogenic MSCs. Taken together, these results suggest that Wnt signaling crosstalk and functional antagonism with the LRP5 co-receptor are key signaling regulators of MSC maintenance and differentiation., ((c) 2007 Wiley-Liss, Inc.)
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- 2007
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15. Comparison of proliferative and multilineage differentiation potential of human mesenchymal stem cells derived from umbilical cord and bone marrow.
- Author
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Baksh D, Yao R, and Tuan RS
- Subjects
- CD146 Antigen metabolism, Cell Lineage, Cells, Cultured, Humans, Immunophenotyping, Mesenchymal Stem Cells metabolism, Signal Transduction, Transfection, Wnt Proteins metabolism, Bone Marrow Cells cytology, Cell Differentiation, Cell Proliferation, Mesenchymal Stem Cells cytology, Umbilical Cord cytology
- Abstract
Human umbilical cord perivascular cells (HUCPVCs) have been shown to have a high proliferative potential and the capacity to differentiate into an osteogenic phenotype. HUCPVCs have thus been considered a possible extra-embryonic mesenchymal stem cell (MSC) source for cell-based therapies. To assess this potential, we compared HUCPVCs to the "gold standard" bone marrow mesenchymal stromal cells (BMSCs) with respect to their proliferation, differentiation, and transfection capacities. HUCPVCs showed a higher proliferative potential than BMSCs and were capable of osteogenic, chondrogenic, and adipogenic differentiation. Interestingly, osteogenic differentiation of HUCPVCs proceeded more rapidly than BMSCs. Additionally, HUCPVCs expressed higher levels of CD146, a putative MSC marker, relative to BMSCs. HUCPVCs showed comparable transfection efficiency as BMSCs using a nucleofection method but were more amenable to transfection with liposomal methods (FuGENE). Gene array analysis showed that HUCPVCs also expressed Wnt signaling pathway genes that have been implicated in the regulation of MSCs. The similar characteristics between HUCPVCs and MSCs support the applicability of HUCPVCs for cell-based therapies. Disclosure of potential conflicts of interest is found at the end of this article.
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- 2007
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16. Soluble factor cross-talk between human bone marrow-derived hematopoietic and mesenchymal cells enhances in vitro CFU-F and CFU-O growth and reveals heterogeneity in the mesenchymal progenitor cell compartment.
- Author
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Baksh D, Davies JE, and Zandstra PW
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- Cell Differentiation, Cell Lineage, Cell Proliferation drug effects, Cells, Cultured, Colony-Forming Units Assay, Culture Media, Serum-Free, Fibroblasts, Hematopoietic Stem Cells metabolism, Humans, Interleukin-3 pharmacology, Interleukin-3 Receptor alpha Subunit, Leukocyte Common Antigens metabolism, Mesenchymal Stem Cells metabolism, Receptors, Interleukin-3 metabolism, Stem Cell Factor pharmacology, Hematopoietic Stem Cells cytology, Mesenchymal Stem Cells cytology, Osteoblasts cytology
- Abstract
The homeostatic adult bone marrow (BM) is a complex tissue wherein physical and biochemical interactions serve to maintain a balance between the hematopoietic and nonhematopoietic compartments. To focus on soluble factor interactions occurring between mesenchymal and hematopoietic cells, a serum-free adhesion-independent culture system was developed that allows manipulation of the growth of both mesenchymal and hematopoietic human BM-derived progenitors and the balance between these compartments. Factorial experiments demonstrated a role for stem cell factor (SCF) and interleukin 3 (IL-3) in the concomitant growth of hematopoietic (CD45+) and nonhematopoietic (CD45-) cells, as well as their derivatives. Kinetic tracking of IL-3alpha receptor (CD123) and SCF receptor (CD117) expression on a sorted CD45- cell population revealed the emergence of CD45-CD123+ cells capable of osteogenesis. Of the total fibroblast colony-forming units (CFU-Fs) and osteoblast colony-forming units (CFU-O), approximately 24% of CFU-Fs and about 22% of CFU-Os were recovered from this population. Cell-sorting experiments demonstrated that the CD45+ cell population secreted soluble factors that positively affect the survival and proliferation of CFU-Fs and CFU-Os generated from the CD45- cells. Together, our results provide insight into the intercellular cytokine network between hematopoietic and mesenchymal cells and provide a strategy to mutually culture both mesenchymal and hematopoietic cells in a defined scalable bioprocess.
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- 2005
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17. Human umbilical cord perivascular (HUCPV) cells: a source of mesenchymal progenitors.
- Author
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Sarugaser R, Lickorish D, Baksh D, Hosseini MM, and Davies JE
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- Cells, Cultured, Colony-Forming Units Assay, Fibroblasts cytology, Fibroblasts physiology, Humans, Mesenchymal Stem Cells cytology, Osteogenesis physiology, Umbilical Cord cytology, Umbilical Cord ultrastructure, Cell Differentiation physiology, Mesenchymal Stem Cells physiology, Umbilical Cord physiology
- Abstract
We describe the isolation of a nonhematopoietic (CD45-, CD34-, SH2+, SH3+, Thy-1+, CD44+) human umbilical cord perivascular (HUCPV) cell population. Each HUCPV cell harvest (2-5 x 10(6), depending on the length of cord available) gave rise to a morphologically homogeneous fibroblastic cell population, which expressed alpha-actin, desmin, vimentin, and 3G5 (a pericyte marker) in culture. We determined the colony-forming unit-fibro-blast (CFU-F) frequency of primary HUCPV cells to be 1:333 and the doubling time, which was 60 hours at passage 0 (P0), decreased to 20 hours at P2. This resulted in a significant cell expansion, producing over 10(10) HUCPV cells within 30 days of culture. Furthermore, HUCPV cells cultured in nonosteogenic conditions contained a subpopulation that exhibited a functional osteogenic phenotype and elaborated bone nodules. The frequency of this CFU-osteogenic subpopulation at P1 was 2.6/10(5) CFU-F, which increased to 7.5/10(5) CFU-F at P2. Addition of osteogenic supplements to the culture medium resulted in these frequencies increasing to 1.2/10(4) and 1.3/10(4) CFU-F, respectively, for P1 and P2. CFU-O were not seen at P0 in either osteogenic or non-osteogenic culture conditions, but P0 HUCPV cells did contain a 20% subpopulation that presented neither class I nor class II cell-surface major histocompatibility complexes (MHC-/-). This population increased to 95% following passage and cryopreservation (P5). We conclude that, due to their rapid doubling time, high frequencies of CFU-F and CFU-O, and high MHC-/- phenotype, HUCPV cells represent a significant source of cells for allogeneic mesenchymal cell-based therapies.
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- 2005
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18. Adult human bone marrow-derived mesenchymal progenitor cells are capable of adhesion-independent survival and expansion.
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Baksh D, Davies JE, and Zandstra PW
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- Adult, Bone Marrow Cells drug effects, Cell Adhesion, Cell Culture Techniques methods, Cell Division drug effects, Cell Survival drug effects, Cells, Cultured cytology, Cells, Cultured drug effects, Cytokines pharmacology, Hematopoiesis, Humans, Middle Aged, Multipotent Stem Cells drug effects, Suspensions, Bone Marrow Cells cytology, Mesoderm cytology, Multipotent Stem Cells cytology
- Abstract
Overview: We show the existence of adult human mesenchymal progenitor cells (hMPCs) that can proliferate, in a cytokine-dependent manner, as individual cells in stirred suspension cultures (SSC) while maintaining their ability to form functional differentiated mesenchymal cell types., Materials and Methods: Ficolled human bone marrow (BM)-derived cells were grown in SSC (and adherent controls) in the presence and absence of exogenously added cytokines. Phenotypic, gene expression, and functional assays for hematopoietic and nonhematopoietic cell populations were used to kinetically track cell production. Limiting-dilution analysis was used to relate culture-produced cells to input cell populations., Results: Cytokine cocktail influenced total and progenitor cell expansion, as well as the types of cells generated upon plating. Flow cytometric analysis of CD117, CD123, and CD45 expression showed that cytokine supplementation influenced SSC output. The concomitant growth of CD45(+) and CD45(-) cells in the cultures that exhibited the greatest hMPC expansions suggests that the growth of these cells may benefit from interactions with hematopoietic cells. Functional assays demonstrated that the SSC-derived cells (input CFU-O number: 1990+/-377) grown in the presence of SCF+IL-3 resulted, after 21 days, in the generation of a significantly greater number (p<0.05) of bone progenitors (33,700+/-8763 CFU-O) than similarly initiated adherent cultures (214+/-75 CFU-O). RT-PCR analysis confirmed that the SSC-derived cells grown in osteogenic conditions express bone-specific genes (Cbfa1/Runx2, bone sialoprotein, and osteocalcin)., Conclusions: Our approach not only provides an alternative strategy to expand adult BM-derived nonhematopoietic progenitor cell numbers in a scalable and controllable bioprocess, but also questions established biological paradigms concerning the properties of connective-tissue stem and progenitor cells.
- Published
- 2003
- Full Text
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