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 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.
['Mung|Kwan-Long|KL|', 'Tsui|Yat-Ping|YP|', 'Tai|Evelyn Wing-Yin|EW|', 'Chan|Ying-Shing|YS|', 'Shum|Daisy Kwok-Yan|DK|', 'Shea|Graham Ka-Hon|GK|']
D000818:Animals D018929:Cell Culture Techniques D002454:Cell Differentiation D002478:Cells, Cultured D018920:Coculture Techniques D016222:Fibroblast Growth Factor 2 D005727:Ganglia, Spinal D006801:Humans D000860:Hypoxia D059630:Mesenchymal Stem Cells D058953:Neural Stem Cells D055495:Neurogenesis D009457:Neuroglia D009474:Neurons D051381:Rats D012583:Schwann Cells D013234:Stem Cells
Bone marrow stromal cell Hypoxia Neural progenitor cell Schwann cell Stem cell niche