【Animal Modeling】-How to establish a mouse testis in vitro culture model?

  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 is used to observe the changes of testicular tissue structure, BrdU staining is used to observe the proliferation of testicular cells in vitro, radioimmunoassay is used to observe the secretion ability of testis in vitro, and the testis is 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 spermatogonia proliferation index of each group showed an upward trend, while the Sertoli cell proliferation index showed a downward trend: the increase of the spermatogonia index of the 3d group was statistically significant compared with that of 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. (Vimentin) expression is seen.

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