动物造模,小鼠睾丸体外培养模型 z-bio 2022-11-24 305

　　Objective: To establish a mouse testis organ culture model by exploring suitable culture conditions.

　　Methods: The testis was cultured in vitro by rotating ventilation culture, and the culture conditions were optimized. The morphology of testis was compared with Transwell cell culture. HE staining was used to observe the changes of testicular tissue structure, BrdU staining was used to observe the cell proliferation of testes in vitro, radioimmunoassay was used to observe the testosterone secretion of testes in vitro, and immunohistochemistry was used to observe the expression of functional proteins in testes.

　　Results: Compared with the results of culture, the morphology of testis tissue cultured by rotating ventilation culture method was more complete. The testis should be 0.3~0.8 mm3 in size. The proliferation index of spermatogonia in each group showed an upward trend, while that of Sertoli cells showed a downward trend: the increase of the proliferation index of spermatogonia in the 3 d group was statistically different from that in the 1 d group (P<0.05), and the decrease of the proliferation index of Sertoli cells in the 5 d group was statistically different from that in the 1 d group (P<0.05). The amount of testosterone secretion decreased with the increase of culture days and the difference was statistically significant (P<0.05). After 3 days of culture, 3 β- Hydroxysteroid Dehydrogenase (3 β- HSD), cytochrome P450 17 α The expression of hydroxylase (P450c17), cholesterol side chain lyase (P450Scc) and vimentin in the cytoplasm of Sertoli cells.

　　Conclusion: The model of testicular organ culture in vitro was successfully established by the method of rotary aeration culture.