The present invention relates to a method for isolating and culturing chicken primordial
germline stem cells (PGCs) and constructing genetically modified chickens. Under an optimized culture
system, short-term
in vitro culture can yield millions of cells and establish stable lines. These lines can also be cultured for over 200 days
in vitro while maintaining migration capacity. After genetic modification of the PGCs established
in vitro, genetically modified chickens can be obtained via vascular injection. Furthermore, PGCs injected into the blood vessels of chicken embryos can be sorted on the seventh day of incubation, recultured in vitro, and then injected back into the blood vessels of the chicken
embryo, thereby improving the efficiency of constructing genetically modified chickens. The present invention also involves freezing PGCs, achieving a
resuscitation survival rate exceeding 80%, allowing for long-term storage of isolated or genetically modified PGCs. The present invention has the advantages of
high isolation and culture efficiency, and the two-vascular injection method can improve the efficiency of constructing genetically modified chickens. The present invention has promising application prospects in the in vitro isolation, culture, line establishment,
cryopreservation, and construction of genetically modified chickens from chicken PGCs.