Department of Biochemistry and Molecular Biology, University of Georgia, Athens, GA, USA.
Nat Protoc. 2013 Jan;8(1):203-12. doi: 10.1038/nprot.2012.156. Epub 2013 Jan 3.
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 ~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.
多能神经嵴干细胞（NCSCs）具有生成多种细胞类型的潜力，包括黑素细胞、外周神经元、平滑肌、骨、软骨和脂肪细胞。本方案详细描述了如何在无饲养层条件下使用化学定义的培养基进行高效、谱系特异性的人类多能细胞向 NCSC 命运分化。该方法使用两种小分子化合物，在约 15 天内实现人类多能细胞向 NCSC 的高效转化。完成本方案后，NCSCs 可用于多种应用，包括生成足够数量的细胞以进行药物筛选、在工业规模上开发细胞治疗药物，并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，从而进一步了解与神经嵴发育相关的人类疾病，例如特雷彻-柯林斯综合征。
