Kumar, U.; Kumar, N.; Shekar, P. C.; Thangaraj, K.

Primordial germ cells (PGCs) are the precursors of germ cells, specified from a subset of epiblast stem cells at the post-implantation stage of mammalian embryonic development. Over the past decade, primordial germ cell-like cells (PGCLCs) have been successfully derived in vitro from mouse and human embryonic stem cells (ESCs). These PGCLCs can further differentiate into mature sperm upon transplantation into neonatal testes. Such advancements offer promising strategies to address male infertility, which affects approximately 15% of couples worldwide. The discovery of induced pluripotent stem cells (iPSCs), which possess similar potential to ESCs, further raises hope for patient-specific therapeutic interventions. However, a major challenge remains: the low efficiency of in vitro PGCLC differentiation. To overcome this, we developed an optimized PGCLC differentiation strategy using a Dppa3-mCherry knock-in reporter mouse ESC line, TDM11. By testing various culture conditions, we identified ascorbic acid as a potent inducer of PGCLC differentiation from ESCs, providing a potential avenue for enhancing in vitro germ cell derivation.