鸡精原细胞分离与纯化初探

Isolation and purification of spermatogonia in chicken

采用二酶三步消化法处理鸡睾丸组织，获得的细胞悬液以Percoll作为介质并结合细胞选择性贴壁培养纯化精原细胞。结果发现：1mg／mL胶原酶＋0．25％胰蛋白酶＋0．25％胰蛋白酶处理鸡睾丸组织，获得的细胞总数多达5．8×10^6个；Percoll离心后精原细胞主要位于19％～35％梯度层中，纯度平均达到66．4％；贴壁纯化后精原细胞纯度则达到82％，比未经贴壁纯化的精原细胞纯度高15．6％。

The study was carried out on isolation and purification of spermotogonia from chicken embryos. After enzymatic digestion of chicken testicular tissues, the single cell suspension were treated by equilibrium density centrifugation in percoll gradients. Then the spermatogonia were further purified by attachment culture in vivo. The results showed that, the treatment of lrng/mL coUeagenase I combined with 0. 25% trypsin was better, and the total number of cells was also higher than 5.82× 10^6 ;the cell bands of 19 - 35 % collect enriched spermatogonia after percoll centrifugation, in which spermatogonia purfication was add to 66.4 % ;82 % of spermatogonia was obtained after further purification by attachment culture.