Highly efficient derivation of pluripotent stem cells from mouse preimplantation and postimplantation embryos in serum-free conditions

Alejandro De Los Angeles, Daiji Okamura, Jun Wu

Research output: Chapter in Book/Report/Conference proceedingChapter

Abstract

Pluripotency refers to the potential of cells to generate all cell types of the embryo proper. Pluripotency spans a spectrum of cellular states. At one polar extreme is naïve pluripotency, which is identified based on the potential to form germline chimeras. At the other polar extreme is primed pluripotency, in which pluripotent cells are primed to differentiate. Mouse naïve PS cells can be derived from preimplantation embryos. Primed epiblast stem (EpiS) cells are typically isolated from epiblasts of early postimplantation mouse embryos. In this chapter, we describe protocols for highly efficient derivation and propagation of murine naïve and primed PS cell lines in serum-free conditions from preimplantation and postimplantation embryos. We describe generation of mouse naïve PS cells using LIF and inhibitors of MEK and GSK3 kinases and of mouse primed PS cells using FGF2 and IWR1 compound which induces the stabilization of Axin proteins.

Original languageEnglish (US)
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages29-36
Number of pages8
DOIs
StatePublished - Jan 1 2019

Publication series

NameMethods in Molecular Biology
Volume2005
ISSN (Print)1064-3745
ISSN (Electronic)1940-6029

Keywords

  • Embryonic stem cells
  • Epiblast stem cells
  • Pluripotency
  • Pluripotent stem cells
  • Postimplantation epiblast
  • Preimplantation epiblast
  • Region-selective epiblast stem cells

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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  • Cite this

    De Los Angeles, A., Okamura, D., & Wu, J. (2019). Highly efficient derivation of pluripotent stem cells from mouse preimplantation and postimplantation embryos in serum-free conditions. In Methods in Molecular Biology (pp. 29-36). (Methods in Molecular Biology; Vol. 2005). Humana Press Inc.. https://doi.org/10.1007/978-1-4939-9524-0_2