Deciphering the heterogeneity of chicken gonadal primordial germ cells via single-cell RNA sequencing.

Chicken primordial germ cells (PGCs) are valuable for breeding, germplasm preservation, and genetic modifications. Here, using single-cell transcriptome analysis, we found that three transcriptionally distinct PGC subtypes were exist in chicken embryonic gonads PGCs. We further demonstrated that in vitro-cultured PGCs could integrate into recipient gonads via blood vessel injection. Additionally, the gene-modified PGCs could be re-established and integrated into recipient gonads after the PGCs were cultured for long-term use by a second blood-vessel injection. Together, these results provide evidence for substantial heterogeneity among chicken gonadal PGCs, which were previously considered a homogeneous cell population. This diversity could be leveraged to establish and expand PGCs for long-term in vitro culture from differentiated gonads.