[Preparation and functional identification of testicular Sertoli cells].
OBJECTIVE
To simplify the method for separation and cultivation of rat testicular Sertoli cells with high viability, quantity and expression efficiency.


METHODS
Testicular Sertoli cells from 2 to 3-week-old male Wistar rats were prepared by digestion with collagenase, trypsin and DNase and cultured together with active lymphocytes to observe their killing effect against lymphocytes. After cell culture for 72 h, the Sertoli cells were morphologically observed by different means and identified with transmission electron microscope. Fas ligand and follicle-stimulating hormone receptor (FSHR) were examined immunohistochemically to identify testicular Sertoli cells. SABC method was used for labeling the Fas ligand on the testicular Sertoli cells.


RESULTS
The viability of the isolated and cultured Sertoli cells was more than 90%, and in in vitro culture, Sertoli cells, which expressed the Fas ligand, could kill the active lymphocytes.


CONCLUSIONS
This method improves the efficiency in acquisition of rat testicular Sertoli cells expressing Fas ligand, which are believed to be a potential donor for co-transplantation with parathyroid cells to offer immune privilege.
Meng
An-qi
AQ
Department of Organ Transplantation, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China. xymaq168@163.com
Yu
Li-xin
LX
Li
Yong-mei
YM
Du
Chuan-fu
CF
Yuan
Yi
Y
Han
Shu-ling
SL
D000818:Animals; D002450:Cell Communication / Q000276:immunology; D002469:Cell Separation / Q000379:methods; D002470:Cell Survival / Q000276:immunology; D002478:Cells, Cultured; D053222:Fas Ligand Protein / Q000378:metabolism*; D007150:Immunohistochemistry; D008214:Lymphocytes / Q000166:cytology / Q000276:immunology; D008297:Male; D046529:Microscopy, Electron, Transmission; D051381:Rats; D017208:Rats, Wistar; D011962:Receptors, FSH / Q000378:metabolism*; D012708:Sertoli Cells / Q000166:cytology / Q000378:metabolism* / Q000648:ultrastructure; D013737:Testis / Q000166:cytology
D053222:Fas Ligand Protein; D011962:Receptors, FSH