| To date, the CsA-contained immunosuppressive regimen have effectively prevented acute rejection following organ transplantation, and improved the results of the short-term of the transplantation. However, there are plenty evidences that long-term CsA therapy results chronic CsA nephrotoxicity and chronic allograft nephropathy. Morbidity and mortality in graft recipients remain high. This led the chronic graft dysfunction at end and questioned its long- term therapy in organ transplantation. Thus, minimal calcineurin inhibitors such as CsA is the tendency of the development of an optimal immunosuppressant regimen.In an attempt to induce donor specific immunologic low-reaction in organ transplantation, a number of protocols have been attempted, and some yield ideal results in rodents. Among these protocols, the donor bone marrow cells (DBMC) infusion was emphasis. But DBMC infusion sometimes resulted severe toxic side effects and to date little is known about its mechanism.In this study, we investigate the synergistic action of donor bone marrow and short-term CsA on prolonging cardiac allograft survival in the rat cardiac transplantation model.Part 1 Establishment and comparison of 2 types of rat heterotopic heart transplantationAim To study the advantage and disadvantage of 2 types of rat heterotopic heart transplantation model.Methods All experimental procedures were performed by one surgeon. The iso-strains transplantation were between BN rat to BN rat and the hetero-transplantation were between Lewis rat to BN rat. The aorta and pulmonary artery of donor's heart with the abdominal aorta and inferior vena cava of the receptor were anastomosed, respectively, in the heterotopic abdominal heart transplantation and with the right common carotid artery and the right external jugular vein of the receptor were anastomosed respectively in the heterotopic cervical heart transplantation. The function of the graft was followed up daily by palpation of heart beats. Pathological examination was performed at various time after surgery. Comparison was made on the operation time, operation success rate, and allografts survival time.Results The two types of operation (the heterotopic cervical heart transplantation and the heterotopic abdominal heart transplantation) were performed 100 times, respectively. Failure in the former group was mainly due to artery constriction and in the latter group was bleeding. Success rates of rat celiac and cervical heart transplantation in the formulate 40 times experiments were 85% (17/20) and 90% (18/20), respectively. The operation times in the two groups were not different (73.85±6.43min vs 70.7±8.67min. p=0.199). The survival times of the iso-allografts were longer than 100 days and not rejection. There are no significantly differences in the degree of acute rejection at various time after surgery on the pathological examination between the 2 types of rat heterotopic allo-heart transplantation.Conclusions Establishment the 2 types of model were easy operation, short duration of the operation and high rate of success and may use in experimental study widely. The selection of rat heart transplantation depends upon the requirement of research. Part 2 Purify major cells from rat bone marrow cells and analyze their immunologic regulatory effectsAim To isolated donor rat bone marrow cells(DBMC), mesenchymal stem cells(MSC) and immature dendritic cells(imDC) from the rat bone marrow and analyzed their immunologic regulatory effects.Methods The rat DBMC, MSC and imDC from rat femora marrow cells were separated and purified by gradient centrifugation and adherence to the culture protocol. The morphological character of the cultured MSC and imDC was observed under light microscope and electron microscope. Expression of CD molecules were detected by flow cytometry. The isolated DBMC, MSC and imDCs were co-cultured with allogenic spleen cells in the mixed lymphocytes reaction, respectively. The spleen cells proliferation was measured by MTT colorimetry.Results The isolated DBMC, MSC and imDCs had the t...
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