Methods for differentiating pluripotent stem cells in dynamic suspension culture

Methods for differentiating pluripotent stem cells to neuroectoderm in dynamic suspension culture using small molecule or protein inhibitors of TGF[beta]/Activin/Nodal signaling and BMP signaling are provided. Also provided are method and protocols for differentiating pluripotent stem cells such as human embryonic stem cells first to neuroectoderm, then further to glial progenitor cells, and further to oligodendrocyte progenitor cells (OPCs), and compositions obtained thereby. The methods of the present disclosure reproducibly produce neuroectoderm progenitor cells by day 7 of the differentiation process, glial progenitor cells by day 21 of the differentiation process and OPCs by day 42 of the differentiation process. 提供了用于使用TGFβ/激活素/Nodal信号传导和BMP信号传导的小分子或蛋白质抑制剂在动态悬浮培养物中将多能干细胞分化为神经外胚层的方法。还提供了用于将多能干细胞例如人胚胎干细胞首先分化为神经外胚层，然后进一步分化为神经胶质祖细胞，并且进一步分化为少突胶质祖细胞(OPC)的方法和方案，以及由此获得的组合物。本公开内容的方法在分化过程的第7天可再现地产生神经外胚层祖细胞，在分化过程的第21天可再现地产生神经胶质祖细胞，和在分化过程的第42天可再现地产生OPC。