Billon Nathalie, Jolicoeur Christine, Raff Martin
Laboratoire "cellules souches et différenciation," UMR 6543 CNRS, centre de biochimie, Faculté des Sciences Valrose, Nice, France.
Methods Mol Biol. 2006;330:15-32. doi: 10.1385/1-59745-036-7:015.
Oligodendrocytes develop from proliferating oligodendrocyte precursor cells (OPCs), which arise in germinal zones, migrate throughout the developing white matter and divide a limited number of times before they terminally differentiate. Thus far, it has been possible to purify OPCs only from the rat optic nerve, but the purified cells cannot be obtained in large enough numbers for conventional biochemical analyses. Moreover, the central nervous system stem cells that give rise to OPCs have not been purified, limiting the ability to study the earliest stages of commitment to the oligodendrocyte lineage. Pluripotent mouse embryonic stem (ES) cells can be propagated indefinitely in culture and induced to differentiate into various cell types. We describe protocols for culture conditions in which neural precursor cells, OPCs, and oligodendrocytes can be efficiently produced from genetically modified ES cells. This strategy should be useful for study of the intracellular and extracellular factors that direct central nervous system stem cells down the oligodendrocyte pathway and influence subsequent oligodendrocyte differentiation. It may also be useful for producing OPCs and oligodendrocytes from human ES cells for cell therapy and drug screening.
少突胶质细胞由增殖的少突胶质前体细胞（OPCs）发育而来，这些前体细胞起源于生发区，迁移至整个发育中的白质，并在终末分化前进行有限次数的分裂。到目前为止，仅能从大鼠视神经中纯化出OPCs，但纯化后的细胞数量不足以进行常规生化分析。此外，产生OPCs的中枢神经系统干细胞尚未得到纯化，这限制了研究少突胶质细胞谱系定向最早阶段的能力。多能小鼠胚胎干细胞（ES细胞）可在培养中无限增殖，并被诱导分化为各种细胞类型。我们描述了从基因修饰的ES细胞高效产生神经前体细胞、OPCs和少突胶质细胞的培养条件方案。该策略对于研究指导中枢神经系统干细胞沿少突胶质细胞途径分化并影响随后少突胶质细胞分化的细胞内和细胞外因子应是有用的。它对于从人ES细胞产生OPCs和少突胶质细胞用于细胞治疗和药物筛选也可能是有用的。
