This invention discloses a method for the isolation, culture, and identification of chicken sexual gonadal cells, belonging to the field of
biotechnology. The invention selects chicken embryos hatched to 18.5 days old, isolates the gonads after PCR sex identification, and uses 0.25%
trypsin-EDTA
digestion followed by
filtration through a 70μm filter to obtain a single-
cell suspension with a
survival rate >90% and free from PGC
contamination. A sex-specific culture
system is established, allowing cells to be stably passaged and maintain a sex-specific
phenotype. Through morphological, qRT-PCR,
immunofluorescence, and
Western blot multidimensional identification, accurate identification of male testicular
Sertoli cells and female ovarian granulosa cells is achieved. This method is simple to operate, has good reproducibility, and produces
high cell viability and purity. It can provide a stable gonadal microenvironment for the
in vitro directed differentiation of chicken primordial germ cells, significantly improving
gamete induction efficiency. It is suitable for research on avian reproductive and developmental mechanisms, transgenic breeding, and
germplasm resource preservation.