بررسي كلوني زايي سلولهاي بنيادي اسپرماتوگوني منجمد - ذوب شده بالغين در سيستم هم كشتي با سلولهاي سرتولي

Colony Formation Efficiency of Frozen-Thawed Adult Mouse Spermatogonia! Stem Cells in Co-culture with Sertoli Cell

Purpose: The basis of spermatogenesis is the spermatogonia! stem cells. These cells are infrequent in adult testis and their proliferation is low. The aim of present study is Proliferating and enhancing of frozen-thawed spermatogonial cells numbers during in vitro culture as a new avenue to restore spermatogenesis in azoospermia subjects. Materials and Methods: Both Sertoli and spermatogonial cells were isolated from adult mouse testes. The identity of the cells was confirmed by analysis of alkaline phosphatase activity and using immunocytochemistry against Oct-4, c-kit and Vimentin of these cells. Isolated spermatogonial cells were cryopreserved and co-cultured with Sertoli cells during three weeks. Assay of the spermatogonial-cell-derived colonies was carried out at the end of the each week. Results: The viability rate of the frozen cells after thawing (68.4±10.2%) was influenced by cryopreservation procedure significantly (P <0.001). Also, co-culture system with Sertoli cells increased the number and diameter of colonies compared with simple culture groups significantly (P < 0.001). Conclusion: we demonstrated that co-culture system with Sertoli cells can increase in vitro colony formation of adult fresh and frozen-thawed spermatogonial cells.