An efficient and economical culture approach for the enrichment of purified oligodendrocyte progenitor cells

少突胶质细胞 祖细胞 祖细胞 生物 计算生物学 神经科学 细胞生物学 计算机科学 干细胞 髓鞘 中枢神经系统
作者
Jianqin Niu,Lingyun Wang,Shubao Liu,Chengren Li,Jiming Kong,Hai‐Ying Shen,Lan Xiao
出处
期刊:Journal of Neuroscience Methods [Elsevier]
卷期号:209 (1): 241-249 被引量:31
标识
DOI:10.1016/j.jneumeth.2012.05.032
摘要

Oligodendrocyte progenitor cell (OPC) culture has provided a powerful approach to mechanistically investigate the proliferation and differentiation of oligodendroglia. However, existing culture methods (including the traditional shake-off method) have limitations, particularly their low productivities. Therefore, we developed a simplified and highly efficient method to produce a large yield of OPCs with low expense by using specialised modified media, in which B104-conditioned medium (B104-CM) instead of growth factors was used as a mitogenic source for OPC propagation, while a modified OPC isolation-medium was applied to improve the isolation of OPCs. First, we withdrew foetal bovine serum when primary mixed glial cultures were 65–75% confluent and substituted with modified oligodendrocyte growth medium to enrich OPCs. Second, we employed a chemical-based method to isolate and purify OPCs from mixed glial cultures using a modified oligodendrocyte isolation medium. As a result, our approach produced a high yield of purified OPCs, approximately 90-fold higher than that produced via the traditional shake-off method. Importantly, the purified OPCs produced via our modified approach maintained typical capacities of proliferation and differentiation observed in oligodendrocyte lineage cells. Together, our modified method provides a highly efficient approach to OPC culture for oligodendrocyte research.

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