Derivation of neural crest cells from human pluripotent stem cells
Derivation of neural crest cells from human pluripotent stem cells
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DOI：
10.1038/nprot.2010.35
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发表时间：
2010-01-01
期刊：
影响因子：
14.8
通讯作者：
Studer, Lorenz
中科院分区：
文献类型：
--
作者：
Lee, Gabsang;Chambers, Stuart M.;Studer, Lorenz
人类多能干细胞（HPSC）衍生的神经rest（NC）细胞提出了建模人类NC发育方面的有价值的工具，包括细胞命运规格，多能力和细胞迁移。 HPSC衍生的NC细胞也适用于对人类疾病进行建模，并作为可再生细胞源用于再生医学的应用。在这里，我们为人类胚胎干细胞（HESC）或人类诱导的多能干细胞（HIPSC）提供了方案，该方案使用MS5共培养系统或基于骨骼药理抑制骨骼的药理培养方法的新型培养方法进入神经外皮和NC细胞中的方案形态发生蛋白和转化生长因子β信号通路。此外，我们提出了使用流式细胞术纯化和传播HPSC-NC细胞的方案，并定义了体外培养条件。我们的协议已在多个独立的HESC和HIPSC线路中进行了验证。使用该协议生成纯化的HPSC-NC前体所需的平均时间为2-5周。
Human pluripotent stem cell (hPSC)-derived neural crest (NC) cells present a valuable tool for modeling aspects of human NC development, including cell fate specification, multipotency and cell migration. hPSC -derived NC cells are also suitable for modeling human disease and as a renewable cell source for applications in regenerative medicine. Here we provide protocols for the step-wise differentiation of human embryonic stem cells (hESCs) or human induced pluripotent stem cells (hiPSCs) into neuroectodermal and NC cells using either the MS5 coculture system or a novel defined culture method based on pharmacological inhibition of bone morphogenetic protein and transforming growth factor-beta signaling pathways. Furthermore, we present protocols for the purification and propagation of hPSC-NC cells using flow cytometry and defined in vitro culture conditions. Our protocol has been validated in multiple independent hESC and hiPSC lines. The average time required for generating purified hPSC-NC precursors using this protocol is 2-5 weeks.