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Rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning

BACKGROUND: Bone marrow stromal cells (BMSCs) are attractive as a source of neural progenitors for ex vivo generation of neurons and glia. Limited numbers of this subpopulation, however, hinder translation into autologous cell-based therapy. Here, we demonstrate rapid and efficient conditioning with...

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Autores principales: Mung, Kwan-Long, Tsui, Yat-Ping, Tai, Evelyn Wing-Yin, Chan, Ying-Shing, Shum, Daisy Kwok-Yan, Shea, Graham Ka-Hon
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5055711/
https://www.ncbi.nlm.nih.gov/pubmed/27717376
http://dx.doi.org/10.1186/s13287-016-0409-x
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author Mung, Kwan-Long
Tsui, Yat-Ping
Tai, Evelyn Wing-Yin
Chan, Ying-Shing
Shum, Daisy Kwok-Yan
Shea, Graham Ka-Hon
author_facet Mung, Kwan-Long
Tsui, Yat-Ping
Tai, Evelyn Wing-Yin
Chan, Ying-Shing
Shum, Daisy Kwok-Yan
Shea, Graham Ka-Hon
author_sort Mung, Kwan-Long
collection PubMed
description BACKGROUND: Bone marrow stromal cells (BMSCs) are attractive as a source of neural progenitors for ex vivo generation of neurons and glia. Limited numbers of this subpopulation, however, hinder translation into autologous cell-based therapy. Here, we demonstrate rapid and efficient conditioning with hypoxia to enrich for these neural progenitor cells prior to further expansion in neurosphere culture. METHOD: Adherent cultures of BMSCs (rat/human) were subjected to 1 % oxygen for 24 h and then subcultured as neurospheres with epidermal growth factor (EGF) and basic fibroblast growth factor supplementation. Neurospheres and cell progeny were monitored immunocytochemically for marker expression. To generate Schwann cell-like cells, neurospheres were plated out and exposed to gliogenic medium. The resulting cells were co-cultured with purified dorsal root ganglia (rat) neurons and then tested for commitment to the Schwann cell fate. Fate-committed Schwann cells were subjected to in vitro myelination assay. RESULTS: Transient hypoxic treatment increased the size and number of neurospheres generated from both rat and human BMSCs. This effect was EGF-dependent and attenuated with the EGF receptor inhibitor erlotinib. Hypoxia did not affect the capacity of neurospheres to generate neuron- or glia-like precursors. Human Schwann cell-like cells generated from hypoxia-treated BMSCs demonstrated expression of S100β /p75 and capacity for myelination in vitro. CONCLUSION: Enhancing the yield of neural progenitor cells with hypoxic preconditioning of BMSCs in vitro but without inherent risks of genetic manipulation provides a platform for upscaling production of neural cell derivatives for clinical application in cell-based therapy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13287-016-0409-x) contains supplementary material, which is available to authorized users.
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spelling pubmed-50557112016-10-19 Rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning Mung, Kwan-Long Tsui, Yat-Ping Tai, Evelyn Wing-Yin Chan, Ying-Shing Shum, Daisy Kwok-Yan Shea, Graham Ka-Hon Stem Cell Res Ther Research BACKGROUND: Bone marrow stromal cells (BMSCs) are attractive as a source of neural progenitors for ex vivo generation of neurons and glia. Limited numbers of this subpopulation, however, hinder translation into autologous cell-based therapy. Here, we demonstrate rapid and efficient conditioning with hypoxia to enrich for these neural progenitor cells prior to further expansion in neurosphere culture. METHOD: Adherent cultures of BMSCs (rat/human) were subjected to 1 % oxygen for 24 h and then subcultured as neurospheres with epidermal growth factor (EGF) and basic fibroblast growth factor supplementation. Neurospheres and cell progeny were monitored immunocytochemically for marker expression. To generate Schwann cell-like cells, neurospheres were plated out and exposed to gliogenic medium. The resulting cells were co-cultured with purified dorsal root ganglia (rat) neurons and then tested for commitment to the Schwann cell fate. Fate-committed Schwann cells were subjected to in vitro myelination assay. RESULTS: Transient hypoxic treatment increased the size and number of neurospheres generated from both rat and human BMSCs. This effect was EGF-dependent and attenuated with the EGF receptor inhibitor erlotinib. Hypoxia did not affect the capacity of neurospheres to generate neuron- or glia-like precursors. Human Schwann cell-like cells generated from hypoxia-treated BMSCs demonstrated expression of S100β /p75 and capacity for myelination in vitro. CONCLUSION: Enhancing the yield of neural progenitor cells with hypoxic preconditioning of BMSCs in vitro but without inherent risks of genetic manipulation provides a platform for upscaling production of neural cell derivatives for clinical application in cell-based therapy. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s13287-016-0409-x) contains supplementary material, which is available to authorized users. BioMed Central 2016-10-07 /pmc/articles/PMC5055711/ /pubmed/27717376 http://dx.doi.org/10.1186/s13287-016-0409-x Text en © The Author(s). 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Mung, Kwan-Long
Tsui, Yat-Ping
Tai, Evelyn Wing-Yin
Chan, Ying-Shing
Shum, Daisy Kwok-Yan
Shea, Graham Ka-Hon
Rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning
title Rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning
title_full Rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning
title_fullStr Rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning
title_full_unstemmed Rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning
title_short Rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning
title_sort rapid and efficient generation of neural progenitors from adult bone marrow stromal cells by hypoxic preconditioning
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5055711/
https://www.ncbi.nlm.nih.gov/pubmed/27717376
http://dx.doi.org/10.1186/s13287-016-0409-x
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