Title: Expanding homogeneous culture of human primordial germ cell-like cells maintaining germline features without serum or feeder layers.

Authors: Kobayashi, Mutsumi; Kobayashi, Misato; Odajima, Junko; Shioda, Keiko; Hwang, Young Sun; Sasaki, Kotaro; Chatterjee, Pranam; Kramme, Christian; Kohman, Richie E; Church, George M; Loehr, Amanda R; Weiss, Robert S; Jüppner, Harald; Gell, Joanna J; Lau, Ching C; Shioda, Toshi

Published In Stem Cell Reports, (2022 Mar 08)

Abstract: In vitro expansion of human primordial germ cell-like cells (hPGCLCs), a pluripotent stem cell-derived PGC model, has proved challenging due to rapid loss of primordial germ cell (PGC)-like identity and limited cell survival/proliferation. Here, we describe long-term culture hPGCLCs (LTC-hPGCLCs), which actively proliferate in a serum-free, feeder-free condition without apparent limit as highly homogeneous diploid cell populations maintaining transcriptomic and epigenomic characteristics of hPGCLCs. Histone proteomics confirmed reduced H3K9me2 and increased H3K27me3 marks in LTC-hPGCLCs compared with induced pluripotent stem cells (iPSCs). LTC-hPGCLCs established from multiple human iPSC clones of both sexes were telomerase positive, senescence-free cells readily passaged with minimal cell death or deviation from the PGC-like identity. LTC-hPGCLCs are capable of differentiating to DAZL-positive M-spermatogonia-like cells in the xenogeneic reconstituted testis (xrTestis) organ culture milieu as well as efficiently producing fully pluripotent embryonic germ cell-like cells in the presence of stem cell factor and fibroblast growth factor 2. Thus, LTC-hPGCLCs provide convenient access to unlimited amounts of high-quality and homogeneous hPGCLCs.

PubMed ID: 35148847

MeSH Terms: No MeSH terms associated with this publication