Protocol of Human Primordial Germ Cell-like Cell Generation from Pluripotent Stem Cells in 2D and 3D Culture Systems

Abstract

Simple Summary Human in vitro gametogenesis is the focus of researchers, as pluripotent stem cell derived human primordial germ cell-like cells (hPGCLCs) could not complete the meiotic division. Therefore, generating early hPGCLCs, which provides an opportunity for further investigations to overcome a meiotic block, is a cue of success in deriving haploid gametes in vitro. Thus, we described a protocol for hPGCLC specification of human pluripotent stem cells (hPSCs) through sequential induction with Activin A for 2 days and BMP4 for 6 days in 2D and 3D culture systems. Induction of hPSCs into hPGCLCs demonstrated expression of early primordial germ cell markers, including PRDM1, NANOS3, DAZL, STELLA, SOX17, SSEA1, and cKIT, on the 8th day of hPGCLC generation.