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

Vivian Lu; Mary T. Doan; Irena J. Roy; Alejandro Torres, Jr.; Michael A. Teitell

STAR Protocols (Sep 2022)

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

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

Affiliations

Vivian Lu: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA 90095, USA
Mary T. Doan: Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA 90095, USA
Irena J. Roy: Developmental and Stem Cell Biology Program, University of California at San Francisco, San Francisco, CA 94143, USA
Alejandro Torres, Jr.: Molecular Biology Institute, University of California at Los Angeles, Los Angeles, CA 90095, USA
Michael A. Teitell: Molecular Biology Institute, University of California at Los Angeles, Los Angeles, CA 90095, USA; Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA 90095, USA; Department of Bioengineering, California NanoSystems Institute, and Broad Center for Regenerative Medicine and Stem Cell Research, University of California at Los Angeles, Los Angeles, CA 90095, USA; Department of Pediatrics, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA 90095, USA; Jonsson Comprehensive Cancer Center, David Geffen School of Medicine, University of California at Los Angeles, Los Angeles, CA 90095, USA; Corresponding author

Journal volume & issue: Vol. 3, no. 3
p. 101568

Abstract

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Summary: 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). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

Published in STAR Protocols

ISSN: 2666-1667 (Online)
Publisher: Elsevier
Country of publisher: United States
LCC subjects: Science: Science (General)
Website: https://star-protocols.cell.com/

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