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

Yukiko Ishikura; Hiroshi Ohta; Masahiro Nagano; Mitinori Saitou

doi:10.1016/j.xpro.2022.101544

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.

Authors

Yukiko Ishikura¹, Hiroshi Ohta², Masahiro Nagano², Mitinori Saitou³

Affiliations

¹ Institute for the Advanced Study of Human Biology (WPI-ASHBi), Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan; Department of Anatomy and Cell Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan. Electronic address: y.ishikura@anat2.med.kyoto-u.ac.jp.
² Institute for the Advanced Study of Human Biology (WPI-ASHBi), Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan; Department of Anatomy and Cell Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan.
³ Institute for the Advanced Study of Human Biology (WPI-ASHBi), Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan; Department of Anatomy and Cell Biology, Graduate School of Medicine, Kyoto University, Yoshida-Konoe-cho, Sakyo-ku, Kyoto 606-8501, Japan; Center for iPS Cell Research and Application (CiRA), Kyoto University, 53 Kawahara-cho, Shogoin, Sakyo-ku, Kyoto 606-8507, Japan. Electronic address: saitou@anat2.med.kyoto-u.ac.jp.

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