Establishment of a rotary aerobic culture system for in vitro culture of mouse testis
10.3969/j.issn.1673-4254.2015.01.13
- VernacularTitle:旋转通气培养法建立小鼠睾丸体外培养模型
- Author:
Yi LIU
1
;
Yanfeng ZHU
;
Zhibin GAO
;
Min LI
;
Lingyu ZHONG
;
Dejiao YIN
;
Yun LI
Author Information
1. 四川大学华西公共卫生学院营养食品卫生与毒理学系
- Keywords:
testis;
organ culture;
testosterone;
cell proliferation;
immunohistochemistry
- From:
Journal of Southern Medical University
2015;(1):66-71
- CountryChina
- Language:Chinese
-
Abstract:
Objective To establish an in vitro model of cultured mouse testis using rotary aerobic culture. Methods Rotary aerobic incubation with optimized culture conditions was used for in vitro culture of mouse testis, and the morphology of the cultured testicular tissues was compared with that cultured in Transwell chambers. The changes in the testicular tissue structure were examined using HE staining, and the cell proliferation was assessed with BrdU staining. Testosterone concentrations in the culture medium were tested with radioimmunoassay and the expression of the functionally related proteins in the testis was detected using immunohistochemistry. Results The testicular tissue cultured by optimized rotary aerobic culture presented with more intact histological structure with the size of the testis ranged from 0.3 to 0.8 mm3. In the two culture systems, the prolifeation index of the spermatogonia increased and that of Sertoli cells decreased with time, and such changes in spermatogonia and Sertoli cell proliferation indices became statistically significant at 3 days (P<0.05) and 5 days (P<0.05) of culture, respectively, as compared with those at 1 day. The concentration of testoerone in the culture media decreased significantly with incubation time (P<0.05). At 3 days of culture, the protein expression of 3β-hydroxysteroid dehydrogenase, cytochrome P450 17α-hydroxylase and cholesterol side-chain cleavage enzyme was detected in Leydig cell cytoplasm and vimentin expression in Sertoli cell cytoplasm. Conclusion An in vitro model of cultured mouse testis has been successfully established using rotary aerobic incubation.
