山羊胎儿睾丸细胞体外分离培养及雄性生殖系干细胞的生长行为

摘要：采用4种不同的处理方法消化山羊胎儿睾丸组织，其中0.1%的胶原酶Ⅰ处理15 min后， 用0.25%胰酶处理5 min，经3次洗涤，睾丸细胞分散较好。用不同的培养体系体外培养，结果显示：原代培养山羊胎儿睾丸细胞培养120 h左右，获得桑椹状雄性生殖系干细胞(mGSCs)集落和单层支持细胞，mGSCs集落呈半悬浮式隆突生长，mGSCs集落和支持细胞分区明显；传代培养显示，在小鼠成纤维细胞饲养层上， mGSCs集落和mGSC单细胞被同质化程度较显著，而含有支持细胞的mGSC单细胞和mGSCs集落被同质化程度较弱；传代mGSCs集落和支持细胞共培养体系中，mGSCs集落正常生长，而且集落中细胞间隙紧密，mGSCs分化较慢。

Separation and In vitro Culture of the Goat Fetal Testicular Cells and the Behavior of Male Germ-line Stem Cells(mGSCs)

Abstract： The seminiferous tubules tissues from goat fetus were treated with 4 methods. The treatment with 0.1% colleagenase for 15 min and then with 0.25% trypsin for 5 min showed ideal cell dispersal degree after centrifugation and washing. The in vitro culture of the cells separated with different medium system showed that mulberry-shaped mGSCs clusters and a single layer of Sertoli cells appeared as original generation in 120 h, the mGSCs clusters developed half-suspendedly and distributed in different locations with the Sertoli cells in the plate; The cells in the margin area of incompletely digested mGSCs clusters and the single cells from them were obviously homogenized after culture for 5 days in fiber cell feeding layer from mice fetus, while same-treated mGSCs clusters co-cultured with Sertoli cells did not displayed clear homogenization in marginal cells of the clusters in the same culture system; And the first generation of mGSCs clusters co-cultured with Sertoli cells formed the same morphologic properties with the original generation of clusters, cells in clusters were tight and mGSCs divided slower than one in MEF feeding layer.