Combination of In Vivo Cryptorchid Testis and In Vitro Co- Culture System to Obtain High Purification and Proliferation of Mouse Spermatogonial Stem Cells

The present study was designed to evaluate the survival and proliferation of spermatogonial stem cells from cryptorchid mouse testis in co-culture system over a 3 weeks period. Sertoli and spermatogonial cells were isolated from bilateral cryptorchid mouse model testes. Isolated spermatogonial cells were co-cultured with Sertoli cells in minimal essential medium (α-MEM) supplemented with 10% fetal calf serum (FCS) for three weeks. The identity of the cells was confirmed through immunocytochemistry against Oct-4 and Vimentin. Best results were achieved from the co-culture system spermatogonia which continued to proliferate, and eventually, type A spermatogonia colonies were found. Most of the cells in these colonies were Oct-4 positive. Bilateral cryptorchid surgery model is a good model for enrichment of spermatogonial stem cells (SSCs). These cells can be used for molecular characterization, genetic manipulation and restoration of male fertility.