绵羊精原干细胞的分离纯化与鉴定

Enrichment and Identification of Spermatogonial Stem Cells from Sheep Testis

旨在探究一种简便高效的绵羊精原干细胞(SSCs)分离纯化方法,为后续研究SSCs奠定基础。本实验采用两步酶消化法对4月龄的绵羊睾丸进行消化处理,获得单细胞悬液。将睾丸细胞接种于0.2%的明胶预处理培养皿中,根据各细胞贴壁能力和速度的不同将SSCs纯化,并在纯化后用SSCs特异标记分子PLZF进行免疫荧光鉴定。结果表明,经免疫荧光染色得出存在PLZF阳性信号。这说明采用两步酶消化和差异贴壁法可以得到纯度较高的SSCs。

The study was aimed to explore a metliod that can eficiently isolate and purify spermatogonial stem cells （ SSCs） from sheep testis,and lay the foundation of the follow - up study of SSCs. Single cellsuspension was prepared from testis of 4 - month old sheep by two - step enzymatic digestion. The SSCswere purified from the suspended cells via differential attachment in 0. 2% gelatin coated dishes,according to the differential adherence capability in different kinds of cells. Immunofluorescence staining by using SSCs molecular marker PLZF was performed after cells were purified. The result shows that the positive signal of PLZF was obtained in the cells after purification. It is indicated that adopting two -step enzymatic digestion and differential attachment can get high purity of SSCs.