成年牛睾丸组织体外无血清培养研究

Study on Serum-free Culture of Adult Bovine Testicular Tissues

为研究促进精子发生的体外培养体系,通过无血清睾丸组织培养法,将成年牛睾丸组织块种植于培养板,观测不同时间睾丸细胞的迁出和形态变化。通过油红O染色鉴定具有分泌功能的支持细胞,通过碱性磷酸酶和免疫组化染色鉴定集落样生长的精原干细胞。结果发现该体系可获得生长良好的多种类型细胞,包括表达AP、Oct-4和GFRα1的精原干细胞及具有分泌功能的睾丸支持细胞,并可获得大量精子。说明,该无血清培养体系可作为一个重要的工具来研究生物学和化学因素对雄性生殖系统的影响,也为研究精子体外发生和移植提供了材料。

In order to study the culture environment of spermatogenesis in vitro, serum-free testicular tissue culture was made. The adult bovine testicular tissue pieces were placed in culture dishes, and movement and morphological changes of tes- ticular cells were observed in different time. Sertoli cells were identified with secretary function by oil red O staining, and sper- matogonial stem cells with colony-like growth were identified by AP and immunohistoehemical staining. A variety of cell types were found in the culture system, including spermatogonial stem cells of AP, Oct-4 and GFRal, sertoli cells with secretary function, and a large number of mature sperms. The results showed that the serum-free culture system could represent a model for investigating the effection of biological and chemical factors on the male reproductive system, and be used as material for the study of spermatogenesis and transplantation.