小鼠精母细胞在体外分化为精子细胞

Differentiation of Mouse Spermatocytes to Spermatids during in vitro Culture

目的:探讨未成熟小鼠生精细胞在体外培养条件下的分化情况。方法:用支持细胞(TM4)条件培养液培养出生后14d的BALB/c小鼠睾丸细胞,对照组采用添加1%胎牛血清的MEM培养液维持细胞存活,在不同时段收获细胞,实时定量RT-PCR分析精子细胞特异性基因Tnp2表达的变化。结果:实时定量RT-PCR显示,在体内,Tnp2的表达随小鼠日龄增长而升高,与精子细胞发生过程同步;在睾丸细胞体外培养过程中,Tnp2的表达在TM4条件培养液培养d5显著升高(P<0.05),而对照组始终无变化。结论:在TM4条件培养液的作用下,小鼠精母细胞在体外分化为精子细胞。

Objective： To investigated the differentiation of immature mouse spermatogenic cells during in vitro culture. Methods： Testicular cells of 14-day-old BALB/c mice were dispersed, and incubated at 34℃ with 5% CO2 in TM4 conditioned medium. In control group, dispersed testicular cells were cultured in MEM medium supplemented with 1% FBS in order to keep cells alive. Cells were harvested at different culture days, and real time RT-PCR was applied to analyze the expression levels of the spermatid specific gene Tnp2. Results： Real time RT- PCR showed, Tnp2 expression increased when the age of mouse increased, which synchronized the differentiation of spermatids in vivo. During in vitro culture, Tnp2 expression inceased significantly in TM4 conditioned medium at d 5 （P〈0.05）, while in control group, Tnp2 expression didn＇t increase all the time. Conclusion： Mouse spermatocytes differentiated to spermatids during in vitro culture in TM4 conditioned medium.