Khazaei Mohamad, Ahuja Christopher S, Fehlings Michael G
Division of Genetics and Development, Krembil Research Institute, Toronto, Ontario, Canada.
Institute of Medical Science, University of Toronto, Toronto, Ontario, Canada.
Curr Protoc Stem Cell Biol. 2017 Aug 14;42:2D.20.1-2D.20.14. doi: 10.1002/cpsc.31.
This unit describes protocols for the efficient generation of oligodendrogenic neural progenitor cells (o-NPCs) from human induced pluripotent stem cells (hiPSCs). Specifically, detailed methods are provided for the maintenance and differentiation of hiPSCs, human induced pluripotent stem cell-derived neural progenitor cells (hiPS-NPCs), and human induced pluripotent stem cell-oligodendrogenic neural progenitor cells (hiPSC-o-NPCs) with the final products being suitable for in vitro experimentation or in vivo transplantation. Throughout, cell exposure to growth factors and patterning morphogens has been optimized for both concentration and timing, based on the literature and empirical experience, resulting in a robust and highly efficient protocol. Using this derivation procedure, it is possible to obtain millions of oligodendrogenic-NPCs within 40 days of initial cell plating which is substantially shorter than other protocols for similar cell types. This protocol has also been optimized to use translationally relevant human iPSCs as the parent cell line. The resultant cells have been extensively characterized both in vitro and in vivo and express key markers of an oligodendrogenic lineage. © 2017 by John Wiley & Sons, Inc.
本单元描述了从人诱导多能干细胞(hiPSC)高效生成少突胶质细胞源性神经祖细胞(o-NPC)的方案。具体而言,提供了关于hiPSC、人诱导多能干细胞源性神经祖细胞(hiPS-NPC)和人诱导多能干细胞少突胶质细胞源性神经祖细胞(hiPSC-o-NPC)的维持和分化的详细方法,最终产物适用于体外实验或体内移植。在整个过程中,根据文献和经验,对细胞暴露于生长因子和模式形态发生素的浓度和时间进行了优化,从而形成了一个稳健且高效的方案。使用这种衍生程序,在初始细胞接种后的40天内可以获得数百万个少突胶质细胞源性NPC,这比用于类似细胞类型的其他方案要短得多。该方案还经过优化,以使用与转化相关的人iPSC作为亲本细胞系。所得细胞已在体外和体内进行了广泛表征,并表达少突胶质细胞谱系的关键标志物。© 2017约翰威立父子公司版权所有