从人诱导多能干细胞生成少突性脊神经祖细胞

Q2 Biochemistry, Genetics and Molecular Biology
Mohamad Khazaei, Christopher S. Ahuja, Michael G. Fehlings
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引用次数: 15

摘要

本单元描述了从人诱导多能干细胞(hiPSCs)高效生成少突胶质神经祖细胞(o- npc)的方法。具体而言,提供了hipsc、人诱导多能干细胞衍生神经祖细胞(hiPS-NPCs)和人诱导多能干细胞-少突神经祖细胞(hiPSC-o-NPCs)的维持和分化的详细方法,最终产品适合体外实验或体内移植。在整个过程中,基于文献和经验经验,细胞暴露于生长因子和模式形态因子的浓度和时间都得到了优化,从而形成了一个强大而高效的方案。使用这种衍生程序,可以在初始细胞镀后40天内获得数百万个少突胶质细胞,这比类似细胞类型的其他方案要短得多。该方案还经过优化,可以使用翻译相关的人类iPSCs作为亲本细胞系。由此产生的细胞在体外和体内都得到了广泛的表征,并表达了一个少突细胞谱系的关键标记。©2017 by John Wiley &儿子,Inc。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Generation of Oligodendrogenic Spinal Neural Progenitor Cells From Human Induced Pluripotent Stem Cells

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.

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来源期刊
Current Protocols in Stem Cell Biology
Current Protocols in Stem Cell Biology Biochemistry, Genetics and Molecular Biology-Cell Biology
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期刊介绍: Published in affiliation with the International Society for Stem Cell Research (ISSCR), Current Protocols in Stem Cell Biology (CPSC) covers the most fundamental protocols and methods in the rapidly growing field of stem cell biology. Updated monthly, CPSC will constantly evolve with thelatest developments and breakthroughs in the field. Drawing on the expertise of leading researchers from around the world, Current Protocols in Stem Cell Biology includes methods and insights that will enhance the progress of global research.
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