Derivation of chicken primordial germ cells using an indirect Co-culture system

Primordial germ cells (PGCs) are promising genetic resources for avian studies including modified animals. However, chicken PGCs are slow to proliferate and gradually lose germline competency after long-term culture, which hinders their application in avian biotechnology. Thus, we developed a robust method for the isolation and rapid propagation of PGCs using an indirect co-culture system. PGCs derived from a pair of embryonic chicken gonads were expanded to 1 × 106 within 2 weeks, and no sex bias was observed in. These PGCs presented high capacity of germline transmission and produced donor-derived offspring after injection into the chicken embryos. This system allows the efficient gene-banking of chicken species and can facilitate the production of chickens bearing a desired phenotype via genomic editing.