Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA 90095, USA.
Developmental and Stem Cell Biology Program, University of California at San Francisco, San Francisco, CA 94143, USA.
STAR Protoc. 2022 Jul 19;3(3):101568. doi: 10.1016/j.xpro.2022.101568. eCollection 2022 Sep 16.
Metabolism regulates cell fates during early mammalian cell differentiation. This protocol describes the steps for directed differentiation of primed human pluripotent stem cells (hPSCs) into the three primary germ lineages-ectoderm, endoderm, and mesoderm-using a chemically defined nutrient-balanced media formulation. Although the transient removal and addition of specific nutrients does not occur during embryonic development, manipulation of nutrients provides an accessible method for evaluating how extracellular and intracellular metabolites help determine hPSC fate. For complete details on the use and execution of this protocol, please refer to Lu et al. (2019) and Lu et al. (2022).
代谢调节早期哺乳动物细胞分化过程中的细胞命运。本方案描述了使用化学定义的营养平衡培养基配方将初始人多能干细胞（hPSC）定向分化为三个主要生殖谱系-外胚层、内胚层和中胚层的步骤。尽管在胚胎发育过程中不会发生特定营养素的短暂去除和添加，但营养素的操纵提供了一种可接近的方法来评估细胞外和细胞内代谢物如何帮助决定 hPSC 命运。有关该方案使用和执行的完整详细信息，请参阅 Lu 等人。（2019 年）和 Lu 等人。（2022 年）。
