Home > Browse Issues > Vol.38 No.7
Advanced Study in Enrichment and Identification of Spermatogonial Stem Cells from Goat Testis
Zhang Qinkai1, Jiang Ying1, Wang Mingming1, Dong Huansheng1, Wei Yujie2, Pan Qingjie1*
1Institute of Animal Reproductive Development and Genetic Engineering, Qingdao Agricultural University, Qingdao 266109, China;
2Shandong Laiyang Animal Husbandry Bureau, Laiyang 265200, China
2Shandong Laiyang Animal Husbandry Bureau, Laiyang 265200, China
Abstract: This study developed an optimized stem cell culture method. With this method goat spermatogonial stem cells (gSSCs) can be efficiently isolated and cultured in vitro for a long time to maintain self-renewal and remain undifferentiated state. Single gSSC suspension was obtained from testis of 3-5 month old goat by twostep enzymatic digestion and differential adhesion. The gSSCs were identified by morphology observation， alkaline phosphatase (AKP) staining and cell immunohistochemistry. The gSSCs were co-cultured with the feeder layer of goat sertoli cells (gSCs)， mouse embryonic fibroblast (MEFs) and laminin. The results showed that gSSC colonies were formed in vitro， and were positive AKP staining. The gene expressions of Oct-4， C-myc， CyclinD1， Ngn3 and TERT were identified by RT-PCR in gSSCs. The protein levels of Oct-4， SSEA-1， α6-integrin were also identified by immunohistochemistry staining. According to statistics of gSSCs colonies， the results showed that the number of colonies on sertoli cell feeder layer was more than on the other two feeder layer， and the difference was significant (P<0.05). The gSSCs were cultured for 3-4 generations as well as 2 months in vitro on sertoli cell feeder layer. All the above showed that the gSSCs can be obtained by the methods of two-step enzyme digestion and differential attachment， and the cells can be well cultured in vitro for proliferation on the feeder layer of sertoli cell.
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