Major Tamara, Powers Ann, Tabar Viviane
Center for Basic and Translational Neuroscience, University of Copenhagen, Denmark.
Icahn Mount Sinai School of Medicine, New York, New York.
Curr Protoc Stem Cell Biol. 2017 Nov 8;39:1H.10.1-1H.10.23. doi: 10.1002/cpsc.17.
Oligodendrocytes are the main myelinating cell of the adult CNS and are vulnerable to injury in diverse disorders such as spinal cord injury, stroke, trauma, pharmacological and radiation toxicity, as well as neuroinflammation. Human pluripotent stem cells are attractive sources of oligodendrocyte lineage cells and provide a promising treatment strategy for exogenous myelin repair through transplantation. This unit describes a protocol for the step-wise differentiation of forebrain late oligodendrocyte progenitor cells (OPCs) from human pluripotent stem cells in defined chemical culture conditions. It involves a stepwise progression of oligodendrocyte progenitors through their known developmental phases, starting with the expression of appropriate transcription factors (Olig2, Nkx2.2), the upregulation of PDGFRA, followed by the appearance of O4-expressing cells, then O1 expression and finally mature myelin-binding protein (MBP) expressing cells. Validation of cell fate is performed by extensive transcriptomal profiling, as well in vitro myelination essays with hESCs derived neuronal cells. Recapitulating forebrain oligodendrocyte development may generate cells more suitable for transplantation strategies for disorders primarily involving the telencephalon.
少突胶质细胞是成体中枢神经系统的主要髓鞘形成细胞，在多种疾病中易受损伤，如脊髓损伤、中风、创伤、药物和辐射毒性以及神经炎症。人多能干细胞是少突胶质细胞系细胞的有吸引力的来源，并为通过移植进行外源性髓鞘修复提供了一种有前景的治疗策略。本单元描述了一种在特定化学培养条件下从人多能干细胞逐步分化为前脑晚期少突胶质前体细胞（OPCs）的方案。它涉及少突胶质前体细胞通过其已知发育阶段的逐步进展，从适当转录因子（Olig2、Nkx2.2）的表达开始，PDGFRA的上调，随后出现表达O4的细胞，然后是O1表达，最后是表达成熟髓鞘结合蛋白（MBP）的细胞。通过广泛的转录组分析以及与hESCs衍生神经元细胞的体外髓鞘形成实验来验证细胞命运。重现前脑少突胶质细胞发育可能会产生更适合用于主要涉及端脑的疾病的移植策略的细胞。
