[The research of xeno-lymphocytes apoptosis induced by Sertoli cells isolated by modified methods in vitro].
OBJECTIVE
To modify the isolation and culture method of Sertoli cells and investigate its' effects on xeno-lymphocytes apoptosis.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Yin
Zhu-zeng
ZZ
Institute of Organ Transplantation, Tongji Hospital of Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430030, China.
Zeng
Meng-hua
MH
Xie
Lin
L
Li
Rong
R
Huang
Ya-bing
YB
Zhu
Min
M
Chen
Gang
G
Chen
Shi
S
D000818:Animals; D017209:Apoptosis; D018929:Cell Culture Techniques / Q000379:methods; D002470:Cell Survival / Q000502:physiology; D002478:Cells, Cultured; D051152:Clusterin / Q000378:metabolism; D018920:Coculture Techniques; D053222:Fas Ligand Protein / Q000378:metabolism; D005434:Flow Cytometry; D007150:Immunohistochemistry; D008214:Lymphocytes / Q000166:cytology* / Q000502:physiology; D008297:Male; D051379:Mice; D008807:Mice, Inbred BALB C; D046529:Microscopy, Electron, Transmission; D051381:Rats; D017207:Rats, Sprague-Dawley; D012708:Sertoli Cells / Q000166:cytology* / Q000378:metabolism / Q000648:ultrastructure; D016212:Transforming Growth Factor beta / Q000378:metabolism
D051152:Clusterin; D053222:Fas Ligand Protein; D016212:Transforming Growth Factor beta