| Kidney transplant is one of the most successful treatment for renal failure patients. However after kidnety transplantation, patients still need some medicine to control acute rejecton. Today immune stress therapy can improve patients'symptom and lessen the pain of the patients, however, These treatments are along with many side flaws such as the side effects on body, too expensive and hard to control the dose of these drugs.Recently we find the sertoli cell strongly express Fas-L which can induce T cell died. We have an idea that if we can use the nature effect of sertoli cell to avoid the transplantation organ's acute rejection?Aim: To investigate the expression of Fas-L on sertoli cell. And culture a stable sertoli cell. After that we can use SD rats and Wisdar rats to build a stable acute renal rejecton modle. According Banff investing system we know the serious these rejecton. So we can plant the sertoli cell and testis organ on the transplant renal's capsule. After 10 days of transplantation. We can use pathological methods to invest the serious of renal rejecton. And we can compare which group has more serious acute renal rejecton.Methods:the sertoli cell was purified and culture in viol for 7 days. Which were identify by electroly microscopy, Feulgen stain. The express of Fas-L were test by immunohistochemistry method. All the culture cells were marked by Brdu marker. After 10 days of plant, the sertoli cell and testis organ on the transplant kidney.Kidney transplantation was performed from Wisdar to SD strain(allogeneic), the donor'arteriae renalis and renal vein were put on the self-make rubber septum and underwent the end to end anastomosis surgeon with the receptor's arteriae renalis and renal vein. After that the donor's bladder valva was inosculated with the receptor's bladder. The serum creatinine level were observed in the 1nd,3nd,5nd and7nd dsys after the surgeons , pathological changes were also observed 4 weeks post-transplantationWe use HE stain, PAS stain, and PASM stain to invest whether the plant group or the normal group have more serious acute renal rejecton.Results: 1. We have developed a convenient method of isolation rat testis Sertoli cells preparation and increased it's purity and viability.Decapsulated and removed testis from 12-day-old rat.then the whole seminiferous tubules were dispersed.The subsequent enzymatic digestion was used with 0.5 % trypsin and 0.1 % collagenase, the resulting tissues were made into cell suspension and subjected to hypotonic shock,after that, the Sertoli cells were cultured at 37℃with 5%C02 in 25cm2 flasks. During the course, observed the culture cells with phase-contrast microscopy daily and identified their morphology. The result showed that, after treated with hypotonic shock, the purity and viability of isolated Sertoli cells reached to 85% % and 84 . 5 % respectively.It's increased the quantity and viability of isolated Sertoli cells greatly.2. Incubation of the SC with BrDU at 15μmol/L and for an optimal length of 48h appeared to achieve the highest LI, both of which exceeded 90%, with the labeling identifiable in five consecutive passages. 3. Rat orthotopic kidney transplantation were performed in 70 cases ,and the successful rate of the model was 90%.4..after 7 days post transplantation we can see the cell plant group has less serious acute renal rejection.Conclusion: When use the end to end anastomosis surgeon on the arteriae renalis and renal vein ,we don't need to block the arteriae aorta and inferior caval vein of the receptor, so we can make less inference on the receptors'systemic circulatory system; the donor's bladder valva was inosculated with the receptor's bladder. can decrease the development of ureter incommodious. This is a reliable, reproducibility and highly achievement ratio model of orthotopic kidney transplantation in rat, and can be used on the basic research of kidney transplantation.Sertoli cell can express Fas-L which can decrease the infiltration of T cell. So we can see the occur of acute rejections are much less in sertoli cell plant group. Maybe we can use this kind of cells to avoid renal rejection.
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