Establishment and Identification of Immortalized Sheep Ovarian Granulosa Cells

This study successfully established a functionally stable immortalized sheep ovarian granulosa cell line using an SV40 T lentiviral vector, which allows for continuous passage up to the 50th generation while maintaining stable cell morphology and fluorescence expression. The transfection efficiency was approximately 90% under optimized conditions (MOI = 10). The immortalized cells exhibited a significantly higher growth rate and an increased proportion of S and G1 phases compared to primary cells. They continued to secrete estradiol, maintaining endocrine function, and did not induce tumors in mice, indicating in vivo safety. This cell model provides a valuable tool for studying animal reproductive function and hormone regulation mechanisms.