| Background:Along with the progress of modern cancer treatment, tumor patient survival has been improved greatly. Ovarian tissue is highly sensitive to chemotherapy/radiation therapy, which can cause female patient’s reproductive endocrinological disorder and even the loss of fertility. It is worth noting that more and more women put off their reproductive age to30to40years old. Young cancer patients’long-term survival and their delayed first childbearing age forced them to confront the risk of premature ovarian failure and infertility. Ovarian tissue xenotransplantation is the important way for fertility preservation. Most researches select mice models as receptors for ovarian tissue heterotransplantation at home and abroad. It’s sure for those rodent models, while with some deficiencies such as far relationship between mice and human, difficult to manage SCID mice receptors, and limitation for growth space of xenografts. There has not been a research report about ovarian tissue xenografted into back muscle of rabbit. Choosing rabbit as xenograft model receptor will enrich the existing xenotransplantation rodent model. Moreover, it provides more choice for young female patients of their fertility preservation.Objective:Investigate the research on fresh human ovarian tissue xenografted into the back muscle of rabbit, and evaluation the effect of application of FTY720combined CsA in maintaining long-term survival of ovarian tissue xenografts.Method:Selecting female New Zealand White rabbits (n=20) as fresh human ovarian tissue heterotransplantation receptors. Twenty ovary castrated rabbits were randomly assigned to seven groups (two castrated rabbits in control group). Ovary tissue xenografts and four lymphoid organs were recovered at the end of research (eight-week post-xenotransplantation). Serum E2level was tested by the technical inspection of radioimmunoassay. Serum IFN-Fand IL-4levels were measured by Enzyme-linked immunosorbent assay. Peripheral blood T cells of CD4+/CD8+were analyzed with flow cytometry assay. Microscopic analysis of recovered xenografts was mesured by H&E. Furthermore, Immunohistochemistry of CD31and Ki-67were used to test follicular growth and neovascularization in the grafts. T cells of sacculus rotundus, Popliteal lymph node, spleen, and thymus gland of all the rabbits were immunohistochemistried by TLR-2+/TLR-4+Results:1The estrus cycle re-established in rabbits receiving xenografts.2Compared with blank, CsA, FTY720groups, combined treatment with cyclosporin A and FTY720improved graft survival, reduced peripheral CD4+and CD8+T cell counts. 3Neovascularisation took place much more in the peripheral zone of the xenograft of combined treatment groups, which were lower in blank and CsA groups. Ki-67positive stained granulosa cells of early-stage follicles and stromal cells in combined treatment groups; nevertheless, only stromal cells were weak positive stained in blank and CsA groups.4TLR-2+/TLR-4+T lymphocytes distributed mainly in the popliteal fossa of lymph node and spleen artery sheath around. TLR-2+/TLR-4+T cells gathered much more in the popliteal fossa of lymph node and spleen artery sheath in combined treatment groups vs. blank and CsA groups.Conclusions:1. A novel human ovarian xenografting model was established by therapeutic regimens of FTY720and CsA;2. Back muscle of rabbit is a optimal site supporting xenografts’follicular growth that reduced ischemia-reperfusion injury post transplantation;3. Combined treatment groups (Group1and Group2) effectively maintained ovarian xenografts for eight weeks, while mono-treatment of CsA or FTY720can’t maintain long-term ovarian tissue survival. |
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