Chen Ying, Balasubramaniyan Veerakumar, Peng Jie, Hurlock Edward C, Tallquist Michelle, Li Jianrong, Lu Q Richard
Department of Developmental Biology and Kent Waldrep Foundation Center for Basic Neuroscience Research on Nerve Growth and Regeneration, University of Texas Southwestern Medical Center, Dallas, Texas 75390, USA.
Nat Protoc. 2007;2(5):1044-51. doi: 10.1038/nprot.2007.149.
The ability to isolate oligodendroglial precursor cells (OPCs) provides a powerful means to characterize their differentiation, properties and potential for myelin repair. Although much knowledge is available for isolation of OPCs from the rat central nervous system, preparation and maintenance of mouse OPCs has been until recently a challenge owing to difficulties in obtaining a sufficient quantity of purified OPCs. Here, we describe protocols to prepare highly enriched rat OPCs and nearly homogenous mouse OPCs. The mouse method generates predominantly OPCs from cortical neural progenitor cells as clonal aggregates called "oligospheres" by taking advantage of molecular genetic tools. Isolated OPCs can be further differentiated into oligodendrocytes. Collectively, we describe simple and efficient methods for the preparation and in vitro maintenance of enriched OPCs from rats and mice. Isolation and culture of a large, homogenous population of rodent OPCs should significantly facilitate studies on OPC lineage progression and their utility in myelin repair after injury.
分离少突胶质前体细胞（OPCs）的能力为表征其分化、特性及髓鞘修复潜力提供了有力手段。尽管已有很多从大鼠中枢神经系统分离OPCs的知识，但直到最近，由于难以获得足量纯化的OPCs，制备和维持小鼠OPCs仍是一项挑战。在此，我们描述了制备高度富集的大鼠OPCs和近乎同质的小鼠OPCs的方案。小鼠制备方法利用分子遗传学工具，从皮质神经祖细胞中主要生成作为克隆聚集体（称为“少突球”）的OPCs。分离出的OPCs可进一步分化为少突胶质细胞。总体而言，我们描述了从大鼠和小鼠制备和体外维持富集OPCs的简单有效方法。分离并培养大量同质的啮齿动物OPCs群体应能显著促进对OPC谱系进展及其在损伤后髓鞘修复中的效用的研究。
