Work

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

Year: 2022

Type: article

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) 12. Lu, V. ∙ Dahan, P. ∙ Ahsan, F.M. ... Mitochondrial metabolism and glutamine are essential for mesoderm differentiation of human pluripotent stem cells Cell Res. 2019; 29 :596-598 Google Scholar and Lu et al. (2022) 14. Lu, V. ∙ Roy, I.J. ∙ Torres, Jr., A. ... Glutamine-dependent signaling controls pluripotent stem cell fate Dev. Cell. 2022; 57 :610-623.e8 Google Scholar .

Source: STAR Protocols

DOI: 10.1016/j.xpro.2022.101568

Authors: Vivian Lu, Mary T. Doan, Irena J. Roy, Alejandro Torres, Michael A. Teitell

Institutions: University of California, Los Angeles, University of California, San Francisco, California NanoSystems Institute, UCLA Jonsson Comprehensive Cancer Center

Language: English

Topic: Pluripotent Stem Cells Research

Subfield: Molecular Biology

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: gold

Estimated APC paid: $1,500