Directed differentiation of human pluripotent cells to neural crest stem cells
Directed differentiation of human pluripotent cells to neural crest stem cells
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DOI：
10.1038/nprot.2012.156
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发表时间：
2013-01-01
期刊：
影响因子：
14.8
通讯作者：
Dalton, Stephen
中科院分区：
文献类型：
--
作者：
Menendez, Laura;Kulik, Michael J.;Dalton, Stephen
多能神经嵴干细胞（NCSC）具有产生广泛的细胞类型的潜力，包括黑素细胞;外周神经元;以及平滑肌、骨、软骨和脂肪细胞。该方案详细描述了如何进行人多能细胞向NCSC命运的高效、谱系特异性分化。该方法在无饲养层条件下使用化学成分确定的培养基，并使用两种小分子化合物在类似于15天的时间内实现人类多能细胞向NCSC的有效转化。在完成该方案后，NCSC可用于许多应用，包括产生足够的细胞数量以进行药物筛选，用于工业规模的细胞治疗剂的开发，以及为人类疾病提供稳健的模型。该方案也可应用于患者来源的诱导多能干细胞，从而用于进一步了解与神经嵴发育相关的人类疾病，例如Treacher-Collins综合征。
Multipotent neural crest stem cells (NCSCs) have the potential to generate a wide range of cell types including melanocytes; peripheral neurons; and smooth muscle, bone, cartilage and fat cells. This protocol describes in detail how to perform a highly efficient, lineage-specific differentiation of human pluripotent cells to a NCSC fate. The approach uses chemically defined media under feeder-free conditions, and it uses two small-molecule compounds to achieve efficient conversion of human pluripotent cells to NCSCs in similar to 15 d. After completion of this protocol, NCSCs can be used for numerous applications, including the generation of sufficient cell numbers to perform drug screens, for the development of cell therapeutics on an industrial scale and to provide a robust model for human disease. This protocol can be also be applied to patient-derived induced pluripotent stem cells and thus used to further the knowledge of human disease associated with neural crest development, for example, Treacher-Collins Syndrome.