睾丸体外培养模型 z-bo 2020-12-01 308

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

　　 Method: Use rotating aeration culture method to culture testis in vitro, optimize culture conditions, and compare morphology with Transwell cell culture method. HE staining was used to observe the changes of testicular tissue structure, BrdU staining was used to observe the cell proliferation of testis in vitro, radioimmunoassay was used to observe the testosterone secretion ability of cultured testes in vitro, and the expression of functional protein in testis was observed by immunohistochemistry.

　　Result: Through the comparison of the culture outcome, the testicular tissue cultured by the rotating ventilation culture method is more complete. The size of testis should be 0.3~0.8 mm3. The proliferation index of spermatogonia in each group showed an upward trend, and the proliferation index of Sertoli cells showed a downward trend: the increase in 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 Sertoli cells in the 5 d group The reduction of proliferation index was statistically different compared with 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 culturing for 3 days, 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P450 17α hydroxylase (P450c17), cholesterol side chain lyase (P450Scc) protein expression can be seen in the cytoplasm of Leydig cells. Sertoli cell Vimentin expression can be seen in the plasma.

　　 Conclusion: The model of in vitro culture of testicular organs was successfully established by the rotation ventilation culture method.