The invention relates to an isolated culture method of functional
Chinese softshell turtle ovarian granular cells. The isolated culture method comprises the following steps: S1, collecting ovarian tissues: collecting
Chinese softshell turtle ovarian tissues; s2, separating and collecting a
granular cell layer; s3, digesting and collecting granular cells; s4,
primary culture of granulosa cells: transferring the collected cells into a
cell culture
bottle containing 4 ml of a complete culture medium for
primary culture, wherein the complete culture medium comprises the following main components: a DMEM culture medium, 15% of fetal calf serum, 1% of
penicillin streptomycin and 1% of an
insulin-
transferrin-
selenium additive; and S5, subculturing the granular cells. The isolated culture method and the culture medium
system provided by the invention are simpler and more time-saving and labor-saving; according to the method, the
aromatase CYP19A1 can be expressed normally, the characteristics of the granular cells are kept, the
aromatase CYP19A1 can be expressed normally, it is indicated that the in-vitro cultured cells can keep the functionality of the cells, and the in-vitro proliferation capacity of the granular cells is improved to a great extent.