ABSTRACT
<p><b>OBJECTIVE</b>To simplify the method for separation and cultivation of rat testicular Sertoli cells with high viability, quantity and expression efficiency.</p><p><b>METHODS</b>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.</p><p><b>RESULTS</b>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.</p><p><b>CONCLUSION</b>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.</p>
Subject(s)
Animals , Male , Rats , Cell Communication , Allergy and Immunology , Cell Separation , Methods , Cell Survival , Allergy and Immunology , Cells, Cultured , Fas Ligand Protein , Metabolism , Immunohistochemistry , Lymphocytes , Cell Biology , Allergy and Immunology , Microscopy, Electron, Transmission , Rats, Wistar , Receptors, FSH , Metabolism , Sertoli Cells , Cell Biology , Metabolism , Testis , Cell BiologyABSTRACT
Objective To investigate the new method of ureter bladder anastomosis in renal transplantation.Methods The ureter was pulled into bladder for 1.2 cm through tunnel at the lateral- top of bladder wall,and the ureter fixed on the bladder wall by 2-3 acus with catgut suture.Results Forty of 42 patients had no complications,and recovered very well,except for 1 cases of necrosis caused by acute rejection and 1 case on urine leakage caused by catheter obstruction from blood clot.Conclu- sion This method is simple,easy to operate,safe and reliable with less complications.