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53. Multifunctionalized hydrogels foster hNSC maturation in 3D cultures and neural regeneration in spinal cord injuries

Author

Marchini, A, Raspa, A, Pugliese, R, El Malek, M, Pastori, V, Lecchi, M, Vescovi, A, Gelain, F, Marchini, Amanda, Raspa, Andrea, Pugliese, Raffaele, El Malek, Marina Abd, Pastori, Valentina, Lecchi, Marzia, Vescovi, Angelo L, Gelain, Fabrizio, Marchini, A, Raspa, A, Pugliese, R, El Malek, M, Pastori, V, Lecchi, M, Vescovi, A, Gelain, F, Marchini, Amanda, Raspa, Andrea, Pugliese, Raffaele, El Malek, Marina Abd, Pastori, Valentina, Lecchi, Marzia, Vescovi, Angelo L, and Gelain, Fabrizio

Abstract

Three-dimensional cell cultures are leading the way to the fabrication of tissue-like constructs useful to developmental biology and pharmaceutical screenings. However, their reproducibility and translational potential have been limited by biomaterial and culture media compositions, as well as cellular sources. We developed a construct comprising synthetic multifunctionalized hydrogels, serum-free media, and densely seeded good manufacturing practice protocol-grade human neural stem cells (hNSC). We tracked hNSC proliferation, differentiation, and maturation into GABAergic, glutamatergic, and cholinergic neurons, showing entangled electrically active neural networks. The neuroregenerative potential of the "engineered tissue" was assessed in spinal cord injuries, where hNSC-derived progenitors and predifferentiated hNSC progeny, embedded in multifunctionalized hydrogels, were implanted. All implants decreased astrogliosis and lowered the immune response, but scaffolds with predifferentiated hNSCs showed higher percentages of neuronal markers, better hNSC engraftment, and improved behavioral recovery. Our hNSC-construct enables the formation of 3D functional neuronal networks in vitro, allowing novel strategies for hNSC therapies in vivo

Published
2019

54. Cancer stem cells from peritumoral tissue of glioblastoma multiforme: the possible missing link between tumor development and progression

Author

Angelucci, Cristiana, primary, D’Alessio, Alessio, additional, Lama, Gina, additional, Binda, Elena, additional, Mangiola, Annunziato, additional, Vescovi, Angelo L., additional, Proietti, Gabriella, additional, Masuelli, Laura, additional, Bei, Roberto, additional, Fazi, Barbara, additional, Ciafrè, Silvia Anna, additional, and Sica, Gigliola, additional

Published
2018
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55. Advances in stem cell therapy for amyotrophic lateral sclerosis

Author

Mazzini, Letizia, Ferrari, Daniela, Andjus, Pavle R., Buzanska, Leonora, Cantello, Roberto, De Marchi, Fabiola, Gelati, Maurizio, Giniatullin, Rashid, Glover, Joel C., Grilli, Mariagrazia, Kozlova, Elena, Maioli, Margherita, Mitrecic, Dinko, Pivoriunas, Augustas, Sanchez-Pernaute, Rosario, Sarnowska, Anna, Vescovi, Angelo L., Mazzini, Letizia, Ferrari, Daniela, Andjus, Pavle R., Buzanska, Leonora, Cantello, Roberto, De Marchi, Fabiola, Gelati, Maurizio, Giniatullin, Rashid, Glover, Joel C., Grilli, Mariagrazia, Kozlova, Elena, Maioli, Margherita, Mitrecic, Dinko, Pivoriunas, Augustas, Sanchez-Pernaute, Rosario, Sarnowska, Anna, and Vescovi, Angelo L.

Abstract

Introduction: Amyotrophic Lateral Sclerosis (ALS) is a progressive, incurable neurodegenerative disease that targets motoneurons. Cell-based therapies have generated widespread interest as a potential therapeutic approach but no conclusive results have yet been reported either from pre-clinical or clinical studies. Areas covered: This is an integrated review of pre-clinical and clinical studies focused on the development of cell-based therapies for ALS. We analyze the biology of stem cell treatments and results obtained from pre-clinical models of ALS and examine the methods and the results obtained to date from clinical trials. We discuss scientific, clinical, and ethical issues and propose some directions for future studies. Expert opinion: While data from individual studies are encouraging, stem-cell-based therapies do not yet represent a satisfactory, reliable clinical option. The field will critically benefit from the introduction of well-designed, randomized and reproducible, powered clinical trials. Comparative studies addressing key issues such as the nature, properties, and number of donor cells, the delivery mode and the selection of proper patient populations that may benefit the most from cell-based therapies are now of the essence. Multidisciplinary networks of experts should be established to empower effective translation of research into the clinic.

Published
2018
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56. Establishment of stable iPS-derived human neural stem cell lines suitable for cell therapies

Author

Rosati, J, Ferrari, D, Altieri, F, Tardivo, S, Ricciolini, C, Fusilli, C, Zalfa, C, Profico, D, Pinos, F, Bernardini, L, Torres, B, Manni, I, Piaggio, G, Binda, E, Copetti, M, Lamorte, G, Mazza, T, Carella, M, Gelati, M, Valente, E, Simeone, A, Vescovi, A, Rosati, Jessica, Ferrari, Daniela, Altieri, Filomena, Tardivo, Silvia, Ricciolini, Claudia, Fusilli, Caterina, Zalfa, Cristina, Profico, Daniela C., Pinos, Francesca, Bernardini, Laura, Torres, Barbara, Manni, Isabella, Piaggio, Giulia, Binda, Elena, Copetti, Massimiliano, Lamorte, Giuseppe, Mazza, Tommaso, Carella, Massimo, Gelati, Maurizio, Valente, Enza Maria, Simeone, Antonio, Vescovi, Angelo L., Rosati, J, Ferrari, D, Altieri, F, Tardivo, S, Ricciolini, C, Fusilli, C, Zalfa, C, Profico, D, Pinos, F, Bernardini, L, Torres, B, Manni, I, Piaggio, G, Binda, E, Copetti, M, Lamorte, G, Mazza, T, Carella, M, Gelati, M, Valente, E, Simeone, A, Vescovi, A, Rosati, Jessica, Ferrari, Daniela, Altieri, Filomena, Tardivo, Silvia, Ricciolini, Claudia, Fusilli, Caterina, Zalfa, Cristina, Profico, Daniela C., Pinos, Francesca, Bernardini, Laura, Torres, Barbara, Manni, Isabella, Piaggio, Giulia, Binda, Elena, Copetti, Massimiliano, Lamorte, Giuseppe, Mazza, Tommaso, Carella, Massimo, Gelati, Maurizio, Valente, Enza Maria, Simeone, Antonio, and Vescovi, Angelo L.

Abstract

Establishing specific cell lineages from human induced pluripotent stem cells (hiPSCs) is vital for cell therapy approaches in regenerative medicine, particularly for neurodegenerative disorders. While neural precursors have been induced from hiPSCs, the establishment of hiPSC-derived human neural stem cells (hiNSCs), with characteristics that match foetal hNSCs and abide by cGMP standards, thus allowing clinical applications, has not been described. We generated hiNSCs by a virus-free technique, whose properties recapitulate those of the clinical-grade hNSCs successfully used in an Amyotrophic Lateral Sclerosis (ALS) phase I clinical trial. Ex vivo, hiNSCs critically depend on exogenous mitogens for stable self-renewal and amplification and spontaneously differentiate into astrocytes, oligodendrocytes and neurons upon their removal. In the brain of immunodeficient mice, hiNSCs engraft and differentiate into neurons and glia, without tumour formation. These findings now warrant the establishment of clinical-grade, autologous and continuous hiNSC lines for clinical trials in neurological diseases such as Huntington’s, Parkinson’s and Alzheimer’s, among others.

Published
2018

57. Advances in stem cell therapy for amyotrophic lateral sclerosis

Author

Mazzini, L, Ferrari, D, Andjus, P, Buzanska, L, Cantello, R, De Marchi, F, Gelati, M, Giniatullin, R, Glover, J, Grilli, M, Kozlova, E, Maioli, M, Mitrečić, D, Pivoriunas, A, Sanchez-Pernaute, R, Sarnowska, A, Vescovi, A, Mazzini, Letizia, Ferrari, Daniela, Andjus, Pavle R, Buzanska, Leonora, Cantello, Roberto, De Marchi, Fabiola, Gelati, Maurizio, Giniatullin, Rashid, Glover, Joel C., Grilli, Mariagrazia, Kozlova, Elena N., Maioli, Margherita, Mitrečić, Dinko, Pivoriunas, Augustas, Sanchez-Pernaute, Rosario, Sarnowska, Anna, Vescovi, Angelo L., Mazzini, L, Ferrari, D, Andjus, P, Buzanska, L, Cantello, R, De Marchi, F, Gelati, M, Giniatullin, R, Glover, J, Grilli, M, Kozlova, E, Maioli, M, Mitrečić, D, Pivoriunas, A, Sanchez-Pernaute, R, Sarnowska, A, Vescovi, A, Mazzini, Letizia, Ferrari, Daniela, Andjus, Pavle R, Buzanska, Leonora, Cantello, Roberto, De Marchi, Fabiola, Gelati, Maurizio, Giniatullin, Rashid, Glover, Joel C., Grilli, Mariagrazia, Kozlova, Elena N., Maioli, Margherita, Mitrečić, Dinko, Pivoriunas, Augustas, Sanchez-Pernaute, Rosario, Sarnowska, Anna, and Vescovi, Angelo L.

Abstract

Introduction: Amyotrophic Lateral Sclerosis (ALS) is a progressive, incurable neurodegenerative disease that targets motoneurons. Cell-based therapies have generated widespread interest as a potential therapeutic approach but no conclusive results have yet been reported either from pre-clinical or clinical studies. Areas covered: This is an integrated review of pre-clinical and clinical studies focused on the development of cell-based therapies for ALS. We analyze the biology of stem cell treatments and results obtained from pre-clinical models of ALS and examine the methods and the results obtained to date from clinical trials. We discuss scientific, clinical, and ethical issues and propose some directions for future studies. Expert opinion: While data from individual studies are encouraging, stem-cell-based therapies do not yet represent a satisfactory, reliable clinical option. The field will critically benefit from the introduction of well-designed, randomized and reproducible, powered clinical trials. Comparative studies addressing key issues such as the nature, properties, and number of donor cells, the delivery mode and the selection of proper patient populations that may benefit the most from cell-based therapies are now of the essence. Multidisciplinary networks of experts should be established to empower effective translation of research into the clinic.

Published
2018

60. Wnt5a Drives an Invasive Phenotype in Human Glioblastoma Stem-like Cells

Author

Binda, Elena, primary, Visioli, Alberto, additional, Giani, Fabrizio, additional, Trivieri, Nadia, additional, Palumbo, Orazio, additional, Restelli, Silvia, additional, Dezi, Fabio, additional, Mazza, Tommaso, additional, Fusilli, Caterina, additional, Legnani, Federico, additional, Carella, Massimo, additional, Di Meco, Francesco, additional, Duggal, Rohit, additional, and Vescovi, Angelo L., additional

Published
2017
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61. Results from Phase I Clinical Trial with Intraspinal Injection of Neural Stem Cells in Amyotrophic Lateral Sclerosis: A Long‐Term Outcome

Author

Mazzini, Letizia, Gelati, Maurizio, Profico, Daniela Celeste, Sorarù, Gianni, Ferrari, Daniela, Copetti, Massimiliano, Muzi, Gianmarco, Ricciolini, Claudia, Carletti, Sandro, Giorgi, Cesare, Spera, Cristina, Frondizi, Domenico, Masiero, Stefano, Stecco, Alessandro, Cisari, Carlo, Bersano, Enrica, De Marchi, Fabiola, Sarnelli, Maria Francesca, Querin, Giorgia, Cantello, Roberto, Petruzzelli, Francesco, Maglione, Annamaria, Zalfa, Cristina, Binda, Elena, Visioli, Alberto, Trombetta, Domenico, Torres, Barbara, Bernardini, Laura, Gaiani, Alessandra, Massara, Maurilio, Paolucci, Silvia, Boulis, Nicholas M., and Vescovi, Angelo L.

Abstract

The main objective of this phase I trial was to assess the feasibility and safety of microtransplanting human neural stem cell (hNSC) lines into the spinal cord of patients with amyotrophic lateral sclerosis (ALS). Eighteen patients with a definite diagnosis of ALS received microinjections of hNSCs into the gray matter tracts of the lumbar or cervical spinal cord. Patients were monitored before and after transplantation by clinical, psychological, neuroradiological, and neurophysiological assessment. For up to 60 months after surgery, none of the patients manifested severe adverse effects or increased disease progression because of the treatment. Eleven patients died, and two underwent tracheotomy as a result of the natural history of the disease. We detected a transitory decrease in progression of ALS Functional Rating Scale Revised, starting within the first month after surgery and up to 4 months after transplantation. Our results show that transplantation of hNSC is a safe procedure that causes no major deleterious effects over the short or long term. This study is the first example of medical transplantation of a highly standardized cell drug product, which can be reproducibly and stably expanded ex vivo, comprising hNSC that are not immortalized, and are derived from the forebrain of the same two donors throughout this entire study as well as across future trials. Our experimental design provides benefits in terms of enhancing both intra‐ and interstudy reproducibility and homogeneity. Given the potential therapeutic effects of the hNSCs, our observations support undertaking future phase II clinical studies in which increased cell dosages are studied in larger cohorts of patients. Stem Cells Translational Medicine2019;8:887&897 In this manuscript we present data from a phase I clinical trial on 18 ALS patients treated with human neural stem cells. Treatment has been performed transplanting cells directly into anterior horn of spinal cord. Results suggested that the procedure is feasible, safe and induce a transitory decline of ALS‐FRS‐R score progression.

Published
2019
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62. Phenotypic variation in Aicardi-Goutières syndrome explained by cell-specific IFN-stimulated gene response and cytokine release

Author

Cuadrado, Eloy, Michailidou, Iliana, Van Bodegraven, Emma J., Jansen, Machiel H., Sluijs, Jacqueline A., Geerts, Dirk, Couraud, Pierre Olivier, Filippis, Lidia De, Vescovi, Angelo L., Kuijpers, Taco W., Hol, Elly M., Cuadrado, Eloy, Michailidou, Iliana, Van Bodegraven, Emma J., Jansen, Machiel H., Sluijs, Jacqueline A., Geerts, Dirk, Couraud, Pierre Olivier, Filippis, Lidia De, Vescovi, Angelo L., Kuijpers, Taco W., and Hol, Elly M.

Published
2015

63. Phenotypic variation in Aicardi-Goutières syndrome explained by cell-specific IFN-stimulated gene response and cytokine release

Author

TN groep Hol, Brain, Cuadrado, Eloy, Michailidou, Iliana, Van Bodegraven, Emma J., Jansen, Machiel H., Sluijs, Jacqueline A., Geerts, Dirk, Couraud, Pierre Olivier, Filippis, Lidia De, Vescovi, Angelo L., Kuijpers, Taco W., Hol, Elly M., TN groep Hol, Brain, Cuadrado, Eloy, Michailidou, Iliana, Van Bodegraven, Emma J., Jansen, Machiel H., Sluijs, Jacqueline A., Geerts, Dirk, Couraud, Pierre Olivier, Filippis, Lidia De, Vescovi, Angelo L., Kuijpers, Taco W., and Hol, Elly M.

Published
2015

64. Phenotypic Variation in Aicardi–Goutières Syndrome Explained by Cell-Specific IFN-Stimulated Gene Response and Cytokine Release

Author

Cuadrado, Eloy, primary, Michailidou, Iliana, additional, van Bodegraven, Emma J., additional, Jansen, Machiel H., additional, Sluijs, Jacqueline A., additional, Geerts, Dirk, additional, Couraud, Pierre-Olivier, additional, De Filippis, Lidia, additional, Vescovi, Angelo L., additional, Kuijpers, Taco W., additional, and Hol, Elly M., additional

Published
2015
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65. Human neural stem cell transplantation in ALS: initial results from a phase I trial

Author

Mazzini, Letizia, primary, Gelati, Maurizio, additional, Profico, Daniela, additional, Sgaravizzi, Giada, additional, Projetti Pensi, Massimo, additional, Muzi, Gianmarco, additional, Ricciolini, Claudia, additional, Rota Nodari, Laura, additional, Carletti, Sandro, additional, Giorgi, Cesare, additional, Spera, Cristina, additional, Domenico, Frondizi, additional, Bersano, Enrica, additional, Petruzzelli, Francesco, additional, Cisari, Carlo, additional, Maglione, Annamaria, additional, Sarnelli, Maria, additional, Stecco, Alessandro, additional, Querin, Giorgia, additional, Masiero, Stefano, additional, Cantello, Roberto, additional, Ferrari, Daniela, additional, Zalfa, Cristina, additional, Binda, Elena, additional, Visioli, Alberto, additional, Trombetta, Domenico, additional, Novelli, Antonio, additional, Torres, Barbara, additional, Bernardini, Laura, additional, Carriero, Alessandro, additional, Prandi, Paolo, additional, Servo, Serena, additional, Cerino, Annalisa, additional, Cima, Valentina, additional, Gaiani, Alessandra, additional, Nasuelli, Nicola, additional, Massara, Maurilio, additional, Glass, Jonathan, additional, Sorarù, Gianni, additional, Boulis, Nicholas M, additional, and Vescovi, Angelo L, additional

Published
2015
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69. Endothelial Cells Create a Stem Cell Niche in Glioblastoma by Providing NOTCH Ligands That Nurture Self-Renewal of Cancer Stem-Like Cells

Author

Zhu, Thant S., primary, Costello, Mark A., additional, Talsma, Caroline E., additional, Flack, Callie G., additional, Crowley, Jessica G., additional, Hamm, Lisa L., additional, He, Xiaobing, additional, Hervey-Jumper, Shawn L., additional, Heth, Jason A., additional, Muraszko, Karin M., additional, DiMeco, Francesco, additional, Vescovi, Angelo L., additional, and Fan, Xing, additional

Published
2011
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72. Glycoproteomic Analysis of Glioblastoma Stem Cell Differentiation

Author

He, Jintang, primary, Liu, Yashu, additional, Zhu, Thant S., additional, Xie, Xiaolei, additional, Costello, Mark A., additional, Talsma, Caroline E., additional, Flack, Callie G., additional, Crowley, Jessica G., additional, DiMeco, Francesco, additional, Vescovi, Angelo L., additional, Fan, Xing, additional, and Lubman, David M., additional

Published
2011
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73. Krüppel-Like Family of Transcription Factor 9, a Differentiation-Associated Transcription Factor, Suppresses Notch1 Signaling and Inhibits Glioblastoma-Initiating Stem Cells

Author

Ying, Mingyao, primary, Sang, Yingying, additional, Li, Yunqing, additional, Guerrero-Cazares, Hugo, additional, Quinones-Hinojosa, Alfredo, additional, Vescovi, Angelo L., additional, Eberhart, Charles G., additional, Xia, Shuli, additional, and Laterra, John, additional

Published
2011
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74. Inorganic mercury changes the fate of murine CNS stem cells

Author

Cedrola, S, Guzzi, G, Ferrari, D, Gritti, A, Vescovi, A, Pendergrass, J, La Porta, C, Cedrola, Sabrina, Guzzi, GianPaolo, Ferrari, Daniela, Gritti, Angela, Vescovi, Angelo L, Pendergrass, James C, La Porta, Caterina A M, Cedrola, S, Guzzi, G, Ferrari, D, Gritti, A, Vescovi, A, Pendergrass, J, La Porta, C, Cedrola, Sabrina, Guzzi, GianPaolo, Ferrari, Daniela, Gritti, Angela, Vescovi, Angelo L, Pendergrass, James C, and La Porta, Caterina A M

Abstract

Stem cells isolated from the central nervous system of both embryonic and adult mice can generate neurons and glia through the activation of different patterns of differentiation in dependence of exposure to appropriate epigenetic signals. On the other hand, environmental conditions might affect the proliferation, migration, and differentiation of these cells. We report here, for the first time, that inorganic mercury affects the proliferative and differentiative capacity of adult neuronal stem cells (ANSCs). Actually, inorganic mercury increases apoptosis in ASNC. Furthermore, in stem cell-derived astrocytes, high levels of the 70 kDa heat shock protein (HSP-70) occur, while the levels of GTP-beta-tubulin activity dramatically decrease. Interestingly, when induced to differentiate, inorganic mercury modifies morphological proprieties of astrocytes, while the neuron population is reduced. These results demonstrate that inorganic mercury produces toxicity in the ANSC-derived neuronal population and affects the biological properties of the glial-derived population

Published
2003

80. NOTCH Pathway Blockade Depletes CD133-Positive Glioblastoma Cells and Inhibits Growth of Tumor Neurospheres and Xenografts

Author

Fan, Xing, primary, Khaki, Leila, additional, Zhu, Thant S., additional, Soules, Mary E., additional, Talsma, Caroline E., additional, Gul, Naheed, additional, Koh, Cheryl, additional, Zhang, Jiangyang, additional, Li, Yue-Ming, additional, Maciaczyk, Jarek, additional, Nikkhah, Guido, additional, DiMeco, Francesco, additional, Piccirillo, Sara, additional, Vescovi, Angelo L., additional, and Eberhart, Charles G., additional

Published
2009
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82. DNER, an Epigenetically Modulated Gene, Regulates Glioblastoma-Derived Neurosphere Cell Differentiation and Tumor Propagation

Author

Sun, Peng, primary, Xia, Shuli, additional, Lal, Bachchu, additional, Eberhart, Charles G., additional, Quinones-Hinojosa, Alfredo, additional, Maciaczyk, Jarek, additional, Matsui, William, additional, DiMeco, Francesco, additional, Piccirillo, Sara M., additional, Vescovi, Angelo L., additional, and Laterra, John, additional

Published
2009
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84. Abnormal DNA Methylation of CD133 in Colorectal and Glioblastoma Tumors

Author

Yi, Joo Mi, primary, Tsai, Hsing-Chen, additional, Glöckner, Sabine C., additional, Lin, Steven, additional, Ohm, Joyce E., additional, Easwaran, Hari, additional, James, C. David, additional, Costello, Joseph F., additional, Riggins, Gregory, additional, Eberhart, Charles G., additional, Laterra, John, additional, Vescovi, Angelo L., additional, Ahuja, Nita, additional, Herman, James G., additional, Schuebel, Kornel E., additional, and Baylin, Stephen B., additional

Published
2008
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85. Efficient In Vitro Labeling of Human Neural Precursor Cells with Superparamagnetic Iron Oxide Particles: Relevance for In Vivo Cell Tracking

Author

Neri, Margherita, primary, Maderna, Claudio, additional, Cavazzin, Chiara, additional, Deidda-Vigoriti, Vivien, additional, Politi, Letterio S., additional, Scotti, Giuseppe, additional, Marzola, Pasquina, additional, Sbarbati, Andrea, additional, Vescovi, Angelo L., additional, and Gritti, Angela, additional

Published
2007
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93. Transcriptome Analysis of Mouse Stem Cells and Early Embryos

Author

Sharov, Alexei A, primary, Piao, Yulan, additional, Matoba, Ryo, additional, Dudekula, Dawood B, additional, Qian, Yong, additional, VanBuren, Vincent, additional, Falco, Geppino, additional, Martin, Patrick R, additional, Stagg, Carole A, additional, Bassey, Uwem C, additional, Wang, Yuxia, additional, Carter, Mark G, additional, Hamatani, Toshio, additional, Aiba, Kazuhiro, additional, Akutsu, Hidenori, additional, Sharova, Lioudmila, additional, Tanaka, Tetsuya S, additional, Kimber, Wendy L, additional, Yoshikawa, Toshiyuki, additional, Jaradat, Saied A, additional, Pantano, Serafino, additional, Nagaraja, Ramaiah, additional, Boheler, Kenneth R, additional, Taub, Dennis, additional, Hodes, Richard J, additional, Longo, Dan L, additional, Schlessinger, David, additional, Keller, Jonathan, additional, Klotz, Emily, additional, Kelsoe, Garnett, additional, Umezawa, Akihiro, additional, Vescovi, Angelo L, additional, Rossant, Janet, additional, Kunath, Tilo, additional, Hogan, Brigid L. M, additional, Curci, Anna, additional, D'Urso, Michele, additional, Kelso, Janet, additional, Hide, Winston, additional, and Ko, Minoru S. H, additional

Published
2003
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95. Clonal Analyses and Cryopreservation of Neural Stem Cell Cultures.

Author

Walker, John M., Weiner, Leslie P., Gritti, Angela, Galli, Rossella, and Vescovi, Angelo L.

Abstract

The discovery of stem cell populations in the adult central nervous system (CNS) that continually produce neurons and glial cells, and the hypothesis that they could contribute to neural plasticity/repair, has opened new and exciting areas of research in basic cell biology and regenerative medicine. The success of these studies relies on understanding the functional features and the normal fate of neural stem cells (NSCs) in vivo as well on the development of in vitro culture conditions enabling isolation, extensive propagation, and rigorous characterization of the "putative" NSCs. The neurosphere assay (NSA) has emerged as a valuable tool for isolating embryonic and adult CNS stem cells and for studying their biology. However, because this assay may select and expand a heterogeneous stem/progenitor cell population, rigorous clonal and serial subcloning analyses are required to detect and document stem cell activity and to unequivocally identify bona fide stem cells. We illustrate and discuss methods for the isolation, propagation, cryopreservation, and functional characterization of NSCs, focusing on the essential issue of their clonogenic capacity. [ABSTRACT FROM AUTHOR]

Published
2008
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96. Adult Neural Stem Cells.

Author

Walker, John M., Weiner, Leslie P., Galli, Rossella, Gritti, Angela, and Vescovi, Angelo L.

Abstract

Neural stem cells (NSCs) have been identified in the mature central nervous system (CNS), and they reside in specific areas. Cultures of NSCs can be successfully established in vitro by exploiting the NeuroSphere assay. This methodology relies on the continuous exposure of neural cells to mitogens such as epidermal growth factor and fibroblast growth factor-2. Under these conditions, only NSCs and highly undifferentiated progenitors proliferate, whereas committed precursors and terminally differentiated cells are eliminated from the culture. The proper application of this method to the cells allows the establishment of long-term expanding stable NSC lines, starting from different neural tissues as the adult rodent CNS and human brain tumor specimens. [ABSTRACT FROM AUTHOR]

Published
2008
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100. Clonal Analyses and Cryopreservation of Neural Stem Cell Cultures.

Author

Walker, John M., Zigova, Tanja, Sanberg, Paul R., Sanchez-Ramos, Juan R., Vescovi, Angelo L., Galli, Rossella, and Gritti, Angela

Abstract

The mature mammalian central nervous system (CNS) has long been considered incapable of significant cell turnover. This view has changed over the last few decades. Recently, the existence of de novo neurogenesis in the adult brain and the presence of stem cells in the mammalian CNS have emerged. The adult brain of both rodents and primates has been shown to embody undifferentiated, mitotically active, precursor cells that are multipotent in nature, and can contribute new, differentiated, neurons and glia to specific regions of the mature brain, such as the olfactory bulb (1-5), the hippocampus (6-8), and the cortex (9-11). While the data clearly suggest the presence of stem cells in the adult CNS in vivo, testing the proliferation, self-renewal, and differentiation capacity of"putative" CNS stem cells relies on the development of methodologies that allow for their isolation and extensive propagation in vitro. [ABSTRACT FROM AUTHOR]

Published
2002
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