Activin A-derived human embryonic stem cells show increased competence to differentiate into primordial germ cell-like cells.

摘要: Protocols for specifying human primordial germ cell-like cells (hPGCLCs) from embryonic stem (hESCs) remain hindered by differences between hESC lines, their derivation methods, and maintenance culture conditions. This poses significant challenges establishing reproducible in vitro models of gametogenesis. Here, we investigated the influence activin A (ActA) during on propensity hESCs to differentiate into PGCLCs. We show that continuous ActA supplementation (from blastocyst until formation post-inner cell mass intermediate [PICMI]) first passage PICMI onwards) is beneficial PGCLCs subsequently. Moreover, comparing isogenic primed naive states prior differentiation, showed conversion 4i-state improves differentiation (TNAP [tissue nonspecific alkaline phosphatase]+/PDPN [podoplanin]+) Those expressed several markers, including TFAP2C (transcription factor AP-2 gamma), SOX17 (SRY-box transcription 17), NANOS3 (nanos C2HC-type zinc finger 3), markers associated with migration, CXCR4 (C-X-C motif chemokine receptor 4), LAMA4 (laminin subunit alpha ITGA6 (integrin 6), CDH4 (cadherin suggesting large numbers obtained may be suitable further more mature cells. Finally, derived presence higher competence hPGCLC, particular if transiently converted 4i-state. Our work provides insights delivers an optimized protocol support efficient derivation.