Aug 2026· Biology· Vol 15, pp. 1436· 0 citations· 33 references
Medicine
TL;DR
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 is described.
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.
The approach provides a valuable model for studying human early embryogenesis, particularly the molecular events surrounding ZGA, and is expected that an individual with ~1 year of experience working with human PSC culture would be able to carry out this protocol.
Md. Abdul Mazid, Yun-Pan Li, Min-Die Zhao et al.· Nature Protocols· 0 citations
This review synthesizes current understanding of PGC specification in mouse and human systems, emphasizing the integration of signaling pathways, transcriptional networks, epigenetic reprogramming, and metabolic regulation.
An integrative map of these metabolic networks is provided to provide insight into the metabolism of hPGCLC development that could help improve methods for fully in vitro differentiation and maturation of hPGCLCs.
Madalena Vaz Santos, B. Schomakers, Marta Llobet Ayala et al.· Biology Open· 0 citations
Ri8CLC induction enables blastoid formation even in the absence of MEK inhibition, TGF-β/Activin/Activin/Nodal inhibition and exogenous LIF, revealing a developmental competence consistent with an early embryonic state and providing a tractable model for studying early human embryogenesis.
Arda Odabas, Sıla Unlu, Eren Ozturk et al.· bioRxiv· 0 citations
This patient-specific iPSC line provides a valuable resource for generating choroid plexus organoids and neurons to investigate disease mechanisms and develop therapeutic strategies to investigate disease mechanisms and develop therapeutic strategies.
Weronika Przybala, Swapnil Gupta, H. Fjerdingstad et al.· bioRxiv· 0 citations
A comprehensive, non-integrating method for generating aneuploid human iPSCs from primary AFCs using episomal plasmid electroporation that yields aneuploid iPSC lines suitable for disease modeling, drug screening, and studies of chromosome biology.
Zi-Fan Song, Jing Wang, Peng Yuan et al.· Journal of Visualized Experi...· 0 citations
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