Optimized protocol to derive germline stem-cell-like cells from mouse pluripotent stem cells

STAR Protoc. 2022 Sep 16;3(3):101544. doi: 10.1016/j.xpro.2022.101544. Epub 2022 Jul 16.

Abstract

Male germ-cell development comprises primordial germ-cell (PGC) development, spermatogonium differentiation, and ensuing spermatogenesis. We present a step-by-step protocol for differentiation of mouse pluripotent stem cells (PSCs) into germline stem-cell-like cells (GSCLCs) via PGC-like cell and spermatogonium-like cell intermediates. The differentiation protocol has higher fidelity than our previous protocol. Upon transplantation into testes in vivo or culture for testis transplants, GSCLCs robustly contribute to spermatogenesis, providing a paradigm for PSC-based reconstitution of mammalian male germ-cell development. For complete details on the use and execution of this protocol, please refer to Ishikura et al. (2021).

Keywords: Cell Differentiation; Developmental biology; Stem Cells.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Differentiation
  • Male
  • Mammals
  • Mice
  • Pluripotent Stem Cells*
  • Spermatogenesis
  • Spermatogonia
  • Testis