快速深低温冻存对新生大鼠卵巢组织免疫原性的影响

Influence of rapid cryopreservation on immunogenicity of ovarian tissue in neonatal rats

目的:探讨快速冻存法冻存新生大鼠卵巢器官的效果,以及冻存对卵巢抗原性的影响。方法:采用快速冻存法冻存出生1 d的鼠(新生鼠)卵巢组织,移植入同系去卵巢成年雌性大鼠的肾被膜下,利用免疫组织化学方法及Western blot检测观察移植物内CD8+和CD4+T淋巴细胞的浸润情况。结果:新生鼠卵巢组织移植后动情周期恢复率(66.67%)低于新鲜移植组(90.51%),但无显著性差异。各移植物内毛细血管丰富;有正常发育阶段的各级卵泡、间质腺及闭锁卵泡;CD8+和CD4+阳性细胞计数值及其蛋白表达明显低于新鲜移植组。结论:快速冻存法可有效地冻存新生鼠的卵巢;冻存降低新生鼠卵巢组织抗原性。

Objective： To investigate the effect of rapid freezing on newborn rats＇ ovaries and to study the influence of cryopreservation on ovarian antigenicity. Methods： Ovaries of neonatal rats （one day old） were preserved by rapidly freezing method. Rapidly thawed ovarian tissues were transplanted underneath the kindey capsules of ovariectomized rats. Immunohistochemistry and Western Blot were used to detect the distribution of CD8^＋ and CD4^＋ T lymphocyte in transplantated tissue. Results： The recovery rate of oestrus cycle in freezing transplanted group （66.67%） was lower than that of fresh transplanted group （90. 51%） （ P 〉 0. 05 ）. The transplanted ovarian tissue contained abundant capillary, follicles of stages, interstitial glands and atresic follicles. The numbers of CD8^＋ and CD4^＋ positive cells and protein expression were obviously lower in freezing group than that in fresh group. Conclusion. Rapid freezing can effectively cryopreserve ovaries of neonatal rats, and the immunogenicity of newborn rats ovarian tissue is reduced after cryopreservation.