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Protocol for germ lineage differentiation of primed human pluripotent stem cells using chemically defined, nutrient-balanced media.

ABSTRACT: 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 in vivo during embryonic development, manipulation of nutrients in vitro 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).

SUBMITTER: Lu V

PROVIDER: S-EPMC9307681 | biostudies-literature | 2022 Sep

REPOSITORIES: biostudies-literature

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Protocol for germ lineage differentiation of primed human pluripotent stem cells using chemically defined, nutrient-balanced media.

Lu Vivian V Doan Mary T MT Roy Irena J IJ Torres Alejandro A Teitell Michael A MA

STAR protocols 20220719 3

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 <i>in vivo</i> during embryonic development, manipulation of nutrients <i>in vitro</i> provides an ...[more]

PMID: 35880122

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