An efficient and economical culture approach for the enrichment of purified oligodendrocyte progenitor cells
An efficient and economical culture approach for the enrichment of purified oligodendrocyte progenitor cells
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一种高效且经济的富集纯化少突胶质细胞祖细胞的培养方法
DOI：
10.1016/j.jneumeth.2012.05.032
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发表时间：
2012-07-30
影响因子：
3
通讯作者：
Xiao, Lan
中科院分区：
文献类型：
--
作者：
Niu, Jianqin;Wang, Lingyun;Xiao, Lan
少突胶质细胞祖细胞（OPC）培养为研究少突胶质细胞的增殖和分化机制提供了有力的手段。然而，现有的培养方法（包括传统的摇瓶法）具有局限性，特别是它们的低生产率。因此，我们开发了一种简单、高效的方法，通过使用专门的改良培养基来生产大量的OPCs，其中B104-条件培养基（B104-CM）代替生长因子用作OPC增殖的有丝分裂源，而改良的OPC分离培养基用于改善OPCs的分离。首先，当原代混合神经胶质细胞培养物达到65-75%融合时，我们取出胎牛血清，并用改良的少突胶质细胞生长培养基替代以富集OPCs。其次，我们采用了一种基于化学的方法，使用改良的少突胶质细胞分离培养基从混合胶质细胞培养物中分离和纯化OPCs。因此，我们的方法产生了高产量的纯化OPC，比通过传统的摇落法产生的高约90倍。重要的是，通过我们改进的方法产生的纯化的OPC保持了在少突胶质细胞谱系细胞中观察到的典型增殖和分化能力。总之，我们的改进方法提供了一个高效的方法，OPC培养少突胶质细胞的研究。(c)2012爱思唯尔有限公司版权所有。
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. (c) 2012 Elsevier B.V. All rights reserved.