动物造模,睾丸体外培养模型 z-bio 2021-12-27 300

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

　　Method: Use the rotating aeration culture method to culture the testis in vitro, optimize the culture conditions, and compare the morphology with the Transwell cell culture method. HE staining was used to observe the structural changes of testis, BrdU staining was used to observe the proliferation of testicular cells in vitro, radioimmunoassay was used to observe the secretion capacity of testis in vitro, and the testis was observed by immunohistochemistry.

　　Result: By comparing the culture results, the morphology of the testicular tissue cultured by the rotating aeration culture method has become more complete. The size of the testis was 0.3-0.8 mm3, the proliferation index of spermatogonia in each group showed an upward trend, while the proliferation index of Sertoli cells showed a downward trend: the increase in the proliferation index of spermatogonia in the 3d group was statistically different from that in the 1d group (P \u003c0 .05)), the Sertoli cell proliferation index was significantly higher in the 5d group. Compared with the first day group, there was a statistically significant difference in the decline (P\u003c0.05). The secretion of testosterone decreased with the increase of culture days, and the difference was statistically significant (P\u003c0.05). After 3 days of culture, 3β-hydroxysteroid dehydrogenase (3β-HSD), cytochrome P45017α hydroxylase (P450c17) and cholesterol side chain lyase (P450Scc) were found in the cytoplasm of Leydig cells and vimentin of supporting cells. ) Protein expression.

　　Conclusion: The in vitro culture model of testicular organs was successfully established by rotating aeration culture.