Efficient induction and isolation of human primordial germ cell-like cells from competent human pluripotent stem cells

Naoko Irie, M. Azim Surani

Research output: Chapter in Book/Report/Conference proceedingChapter

19 Citations (Scopus)

Abstract

We recently reported a robust and defined culture system for the specification of human primordial germ cell-like cells (hPGCLCs) from human pluripotent stem cells (hPSCs), both embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) in vitro (Irie et al. Cell 160: 253-268, 2015). Similar attempts previously produced hPGCLCs from hPSCs at a very low efficiency, and the resulting cells were not fully characterized. A key step, which facilitated efficient hPGCLC specification from hPSCs, was the induction of a “competent” state for PGC fate via the medium containing a cocktail of four inhibitors. The competency of hPSCs can be maintained indefinitely and interchangeably with the conventional/low-competent hPSCs. Specification of hPGCLC occurs following sequential expression of key germ cell fate regulators, notably SOX17 and BLIMP1, as well as initiation of epigenetic resetting over 5 days. The hPGCLCs can be isolated using specific cell surface markers without the need for generating germ cell-specific reporter hPSC lines. This powerful method for the induction and isolation of hPGCLCs can be applied to both hESCs and iPSCs, which can be used for advances in human germ line biology.

Original languageEnglish
Title of host publicationMethods in Molecular Biology
PublisherHumana Press Inc.
Pages217-226
Number of pages10
DOIs
Publication statusPublished - 2017
Externally publishedYes

Publication series

NameMethods in Molecular Biology
Volume1463
ISSN (Print)1064-3745

Keywords

  • Embryonic stem cells
  • Epiblasts
  • Epigenetic resetting
  • Germ cell specification
  • Human pluripotent stem cells
  • Human primordial germ cells
  • Induced pluripotent stem cells
  • Primordial germ cell-like cells

ASJC Scopus subject areas

  • Molecular Biology
  • Genetics

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