小鼠生精细胞体外长期培养及超微结构分析

Long-term Culture of Spermatogenic Cells from Mice and Analysis of Ultra-microscopical Structure

目的初步探讨新生小鼠睾丸组织中生精细胞在体外培养条件下的增殖分化条件,建立有效的生精细胞体外成熟模型。方法用酶法和Percoll不连续密度梯度法分离纯化新生小鼠睾丸生精细胞,获取富精原细胞层,采用自制小鼠睾丸组织培养液对分离纯化后的细胞进行体外培养。用电子显微镜观察培养的细胞超微结构。结果新生小鼠生精细胞在用成年小鼠睾丸组织提取液配制的培养基中存活达194d。电子显微镜观察可见,培养后的细胞中含有精子细胞顶体样结构。结论采用小鼠睾丸组织制备的培养液培养的小鼠生精细胞可体外长期培养并显示出一定的分化趋势。

Objective A preliminary study was made on the proliferation and differentiation condition of spermatogenic cells from neonatal mouse testis in order to establish an effective in - vitro developmental model of spermatogenic cells. Methods Spermatogenic cells from neonatal mice testis were isolated by enzymatic digestion and percoll gradient centrifugation, the spermatogonia - rich layer was collected and a long - term culture with a self - made testis tissue culture medium was performed. After 194 days, cells were collected and their ultra - microscopical structure were analyzed by electron microscope, Results Spermatogenic cells from neonatal mice testis could survive for 194 days in the culture medium extracted from adult mice testis. The acrosome - like structure in cultured cells was observed by electron microscopic, Conclusion In the experimental culture condition, mice spermatogenic cells can be cultured for a long time and show a differentiation tendency,