人睾丸圆形精子细胞在体外向长形精子细胞分化

In-vitro Differentiation of Human Testicular Round Spermatids to Elongating Spermatids

目的:探讨人睾丸生精细胞在体外培养条件下发生的分化。方法:取梗阻性无精子症患者睾丸活检标本,机械法离散睾丸细胞。①睾丸细胞混合培养,每24 h观察,计数分析长形精子细胞比率的变化;②显微操作挑取圆形精子细胞,观察单个细胞在微滴培养过程中变形分化的情况。结果:在添加卵泡刺激素、睾酮的HTF培养液中进行睾丸细胞混合培养,24 h后长形精子细胞比率较培养前显著增加(P<0.05);单个圆形精子细胞培养,可见到圆形精子细胞长出鞭毛转化为长形精子细胞,48 h的转化率为3.54%。Vero细胞条件培养液中人睾丸生精细胞的分化与HTF培养液中相同,两者无显著差异。结论:人睾丸组织的圆形精子细胞在体外可分化为长形精子细胞,Vero细胞条件培养液对此分化无促进作用。

Objective： To investigate the differentiation of human testicular spermatogenic cells during in vitro culture. Methods： Testicular cells of obstructive azoospermic patients＇testis biopsies were dispersed employing mechanic methods. Then, ① mixed testicular cells were applied to in vitro culture, and changes of the ratio of elongating spermatids and all round cells were analyzed during mixed cell culture; ② round spermatids were picked up from the mixed cells employing micromanipulator, followed by differentiation of the isolated round spermatids during microdrup culture. Results ： The ratio of the elongating spermatids increased significantly （ P 〈 0.05 ） after 24 hours of mixed cell culture in HTF medium supplemented with FSH and testosterone. During single round spermatid culture, transformation of the round spermatid to elongating spermatid with newly formed flagellum was observed, and the transformation ratio within 48 hours of microdrop culture was 3.54%. The differentiation of human testicular spermatogenic cells cultured in Vero cell conditioned medium was similar to that cultured in HTF medium. Conclusion： Human testicular round spermatids can differentiate to elongating spermatids during in vitro culture. Vero cell conditioned medium does not promote the differentiation of human testicular round spermatids to elongating spermatids.