The research of xeno-lymphocytes apoptosis induced by Sertoli cells isolated by modified methods in vitro.
- Author:
Zhu-zeng YIN
1
;
Meng-hua ZENG
;
Lin XIE
;
Rong LI
;
Ya-bing HUANG
;
Min ZHU
;
Gang CHEN
;
Shi CHEN
Author Information
- Publication Type:Journal Article
- MeSH: Animals; Apoptosis; Cell Culture Techniques; methods; Cell Survival; physiology; Cells, Cultured; Clusterin; metabolism; Coculture Techniques; Fas Ligand Protein; metabolism; Flow Cytometry; Immunohistochemistry; Lymphocytes; cytology; physiology; Male; Mice; Mice, Inbred BALB C; Microscopy, Electron, Transmission; Rats; Rats, Sprague-Dawley; Sertoli Cells; cytology; metabolism; ultrastructure; Transforming Growth Factor beta; metabolism
- From: Chinese Journal of Surgery 2007;45(5):331-334
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo modify the isolation and culture method of Sertoli cells and investigate its' effects on xeno-lymphocytes apoptosis.
METHODSSertoli 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.
RESULTSIn 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.
CONCLUSIONSIt 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.
