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Differentiation of rat bone marrow mesenchymal stem cells into nerve-like cells through co-culture with retinal cells and application of basic fibroblast growth factor in vitro |
LI Qun-Xiu,LIU Xue-Zheng,ZHANG Ke-jian,HOU Yang |
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Abstract Objective To investigate an effective way to differentiate bone marrow mesenchymal stem cells (BMSC) into nerve-like cells and address the low percentage of differentiation and survival of BMSC induced in vitro. Methods BMSC was separated using density gradient centrifugation and adherence screening methods. The surface antigen of BMSC was identified by immunohistochemistry methods using CD31, CD44, CD45, CD105 monoclonal antibody. The experimental groups were divided as follows: a retinal cell plus basic fibroblast growth factor (bFGF) group, bFGF group, and a negative control group. The expression of Neuronal Class Ⅲβ -Tubulin (Tuj1), neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) of induced BMSC was observed on day 3, 7 and 14 by immunocytochemistry. Animation of induced BMSC was observed by the MTT method. Results The morphologic shape of induced BMSC changed after 12 hours and gradually became typical nerve-like cells. Tuj1, NSE and GFAP positive cells could be found after three days by immunocytochemistry. The percentage of these positive cells increased in the total population with induced time. The proliferative ability of the induced BMSC had not been affected markedly. Conclusion An analogical retinal microenvoirenment and bFGF can induce BMSC to differentiate into nerve-like cells and survive in vitro.
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Received: 04 December 2009
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