Production and Analysis of Human Primordial Germ Cell-Like Cells

• Published in: Methods in molecular biology (Clifton, N.J.) Vol. 2195; p. 125
• Main Authors: Mitsunaga, Shino, Shioda, Keiko, Hanna, Jacob H, Isselbacher, Kurt J, Shioda, Toshi
• Format: Journal Article
• Language: English
• Published: United States 2021
• Subjects: Biomarkers; Cell Culture Techniques - methods; Cell Differentiation; Cell Self Renewal; Cells, Cultured; Embryoid Bodies - cytology; Gastrula - cytology; Germ Cells - cytology; Germ Cells - metabolism; Germ Layers - cytology; Humans; Immunohistochemistry - methods; Immunophenotyping - methods; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Primordial germ cell–like cells; Induced pluripotent stem cells; Spin-EB; Naive pluripotency; AggreWell; Primordial germ cells; Embryoid bodies; Primed pluripotency
• ISSN: 1940-6029
• DOI: 10.1007/978-1-0716-0860-9_10

Primordial germ cells (PGCs) are common ancestors of all germline cells. In mammals, PGCs emerge in early-stage embryos around the timing of gastrulation at or near epiblast, and specification of PGCs from their precursor cells involves multiple growth factors secreted by adjacent cells. Recent advancements in germline stem cell biology have made it possible to generate PGC-like cell culture models (PGCLCs for PGC-like cells) from human and mouse pluripotent stem cells by mimicking the embryonic growth factor environment in vitro. Here we describe a method of producing human PGCLCs from primed-pluripotency induced pluripotent stem cells (iPSCs) via temporal conversion to naive pluripotency followed by formation of embryoid bodies (EBs) using the spin-EB method.