A
Chinese softshell turtle ovarian
granular cell separation and culture method comprises separation and culture, the separation comprises the following steps: selecting a female
Chinese softshell turtle individual in a positive spawning period, anesthetizing the female
Chinese softshell turtle individual, killing the female Chinese softshell turtle individual after neck breaking, separating ovarian tissues, cleaning, removing redundant parts, placing the female Chinese softshell turtle individual in a basal culture medium, separating follicles in II-III periods, keeping the integrity of the follicles, performing
enzyme digestion treatment, and culturing the follicles in the basal culture medium. And centrifuging to remove the supernatant to obtain a precipitate containing egg granules with the
diameter of 0.1-0.5 cm, resuspending the precipitate by adopting a
mixed culture medium, and inoculating and culturing. The ovarian granulosa cells separated by the
separation method provided by the invention are high in yield, the method for culturing the ovarian granulosa cells in a specific
mixed culture medium is diversified in alternative factors and small in added factor amount, meanwhile, a NaHCO3-free culture medium is used, the use of CO2 is reduced, the obtained ovarian granulosa cells are kept consistent with the internal environment of an
organism, the cost is reduced, and the method is suitable for large-scale popularization and application. And the in-vitro lifetime of the ovarian granular cells is prolonged.