The research of xeno-lymphocytes apoptosis induced by Sertoli cells isolated by modified methods in vitro / 中华外科杂志

ABSTRACT

<p><b>OBJECTIVE</b>To modify the isolation and culture method of Sertoli cells and investigate its' effects on xeno-lymphocytes apoptosis.</p><p><b>METHODS</b>Sertoli cells which was isolated from 2 - 4 week-old Sprague Dawley (SD) rats, were successfully prepared by collagenase type V, trypsin and DNase I and then identified by electron microscope. Viability and apoptosis of cultured cells were measured by flow cytometry. The apoptosis rates of Balb/c mouse lymphocytes were examined which were co-cultured with Sertoli cells of SD rats by flow cytometry, too. The expression of FasL, TGF-beta(1) and clusterin on Sertoli cells were detected by immunocytochemistry.</p><p><b>RESULTS</b>In the co-cultured system, Sertoli cells accounted for more than 90%. The viability of Sertoli cells was above 95% and the apoptosis rate was 10.87% +/- 3.87% in this study. The lymphocytes apoptosis ratio was 15.52% +/- 0.17% (P < 0.01). Streptavidin-biotin-peroxidase-complex immunochemistry staining showed that the Sertoli cells could express FasL, TGF-beta(1) and clusterin, respectively.</p><p><b>CONCLUSIONS</b>It indicates that the expression of FasL, TGF-beta(1) on the Sertoli cells might relate to the immune privilege, and it supposed to be benefit for xenotransplantation.</p>