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Purification of Immature Neuronal Cells from Neural Stem Cell Progeny
Large-scale proliferation and multi-lineage differentiation capabilities make neural stem cells (NSCs) a promising renewable source of cells for therapeutic applications. However, the practical application for neuronal cell replacement is limited by heterogeneity of NSC progeny, relatively low yield...
Autores principales: | , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Public Library of Science
2011
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3109004/ https://www.ncbi.nlm.nih.gov/pubmed/21687800 http://dx.doi.org/10.1371/journal.pone.0020941 |
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author | Azari, Hassan Osborne, Geoffrey W. Yasuda, Takahiro Golmohammadi, Mohammad G. Rahman, Maryam Deleyrolle, Loic P. Esfandiari, Ebrahim Adams, David J. Scheffler, Bjorn Steindler, Dennis A. Reynolds, Brent A. |
author_facet | Azari, Hassan Osborne, Geoffrey W. Yasuda, Takahiro Golmohammadi, Mohammad G. Rahman, Maryam Deleyrolle, Loic P. Esfandiari, Ebrahim Adams, David J. Scheffler, Bjorn Steindler, Dennis A. Reynolds, Brent A. |
author_sort | Azari, Hassan |
collection | PubMed |
description | Large-scale proliferation and multi-lineage differentiation capabilities make neural stem cells (NSCs) a promising renewable source of cells for therapeutic applications. However, the practical application for neuronal cell replacement is limited by heterogeneity of NSC progeny, relatively low yield of neurons, predominance of astrocytes, poor survival of donor cells following transplantation and the potential for uncontrolled proliferation of precursor cells. To address these impediments, we have developed a method for the generation of highly enriched immature neurons from murine NSC progeny. Adaptation of the standard differentiation procedure in concert with flow cytometry selection, using scattered light and positive fluorescent light selection based on cell surface antibody binding, provided a near pure (97%) immature neuron population. Using the purified neurons, we screened a panel of growth factors and found that bone morphogenetic protein-4 (BMP-4) demonstrated a strong survival effect on the cells in vitro, and enhanced their functional maturity. This effect was maintained following transplantation into the adult mouse striatum where we observed a 2-fold increase in the survival of the implanted cells and a 3-fold increase in NeuN expression. Additionally, based on the neural-colony forming cell assay (N-CFCA), we noted a 64 fold reduction of the bona fide NSC frequency in neuronal cell population and that implanted donor cells showed no signs of excessive or uncontrolled proliferation. The ability to provide defined neural cell populations from renewable sources such as NSC may find application for cell replacement therapies in the central nervous system. |
format | Online Article Text |
id | pubmed-3109004 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2011 |
publisher | Public Library of Science |
record_format | MEDLINE/PubMed |
spelling | pubmed-31090042011-06-16 Purification of Immature Neuronal Cells from Neural Stem Cell Progeny Azari, Hassan Osborne, Geoffrey W. Yasuda, Takahiro Golmohammadi, Mohammad G. Rahman, Maryam Deleyrolle, Loic P. Esfandiari, Ebrahim Adams, David J. Scheffler, Bjorn Steindler, Dennis A. Reynolds, Brent A. PLoS One Research Article Large-scale proliferation and multi-lineage differentiation capabilities make neural stem cells (NSCs) a promising renewable source of cells for therapeutic applications. However, the practical application for neuronal cell replacement is limited by heterogeneity of NSC progeny, relatively low yield of neurons, predominance of astrocytes, poor survival of donor cells following transplantation and the potential for uncontrolled proliferation of precursor cells. To address these impediments, we have developed a method for the generation of highly enriched immature neurons from murine NSC progeny. Adaptation of the standard differentiation procedure in concert with flow cytometry selection, using scattered light and positive fluorescent light selection based on cell surface antibody binding, provided a near pure (97%) immature neuron population. Using the purified neurons, we screened a panel of growth factors and found that bone morphogenetic protein-4 (BMP-4) demonstrated a strong survival effect on the cells in vitro, and enhanced their functional maturity. This effect was maintained following transplantation into the adult mouse striatum where we observed a 2-fold increase in the survival of the implanted cells and a 3-fold increase in NeuN expression. Additionally, based on the neural-colony forming cell assay (N-CFCA), we noted a 64 fold reduction of the bona fide NSC frequency in neuronal cell population and that implanted donor cells showed no signs of excessive or uncontrolled proliferation. The ability to provide defined neural cell populations from renewable sources such as NSC may find application for cell replacement therapies in the central nervous system. Public Library of Science 2011-06-03 /pmc/articles/PMC3109004/ /pubmed/21687800 http://dx.doi.org/10.1371/journal.pone.0020941 Text en Azari et al. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are properly credited. |
spellingShingle | Research Article Azari, Hassan Osborne, Geoffrey W. Yasuda, Takahiro Golmohammadi, Mohammad G. Rahman, Maryam Deleyrolle, Loic P. Esfandiari, Ebrahim Adams, David J. Scheffler, Bjorn Steindler, Dennis A. Reynolds, Brent A. Purification of Immature Neuronal Cells from Neural Stem Cell Progeny |
title | Purification of Immature Neuronal Cells from Neural Stem Cell Progeny |
title_full | Purification of Immature Neuronal Cells from Neural Stem Cell Progeny |
title_fullStr | Purification of Immature Neuronal Cells from Neural Stem Cell Progeny |
title_full_unstemmed | Purification of Immature Neuronal Cells from Neural Stem Cell Progeny |
title_short | Purification of Immature Neuronal Cells from Neural Stem Cell Progeny |
title_sort | purification of immature neuronal cells from neural stem cell progeny |
topic | Research Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3109004/ https://www.ncbi.nlm.nih.gov/pubmed/21687800 http://dx.doi.org/10.1371/journal.pone.0020941 |
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