Comparison between Slow Freezing and Vitrification for Human Ovarian Tissue Cryopreservation and Xenotransplantation
Comparison between Slow Freezing and Vitrification for Human Ovarian Tissue Cryopreservation and Xenotransplantation
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DOI：
10.3390/ijms20133346
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发表时间：
2019-07-01
影响因子：
5.6
通讯作者：
Kim, Tak
中科院分区：
文献类型：
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作者：
Lee, Sanghoon;Ryu, Ki-Jin;Kim, Tak
利用异种移植模型对两种人类卵巢组织冷冻保存方法进行比较，以建立安全有效的冷冻保存方法。卵巢组织取自在大学医院妇科研究室接受良性卵巢手术的女性。缓慢冷冻或玻璃化冷冻保存4周后，将组织移植到112只切除卵巢的雌性严重联合免疫缺陷小鼠体内。 4周后取回组织。冷冻保存和异种移植后原始卵泡计数减少，且慢速冷冻组显着高于玻璃化组（p < 0.001）。免疫组织化学和TUNEL检测显示，慢速冷冻组的卵泡增殖和血管生成的Ki-67和CD31标记物较高（分别为p < 0.001和p = 0.006），而玻璃化组的DNA损伤更大（p < 0.001）。蛋白质印迹显示玻璃化冷冻增加了细胞凋亡。在接受两种冷冻保存技术的移植样本中，抗苗勒氏管激素表达较低。电子显微镜显示玻璃化冷冻组的原始卵泡变形。就异种移植后卵泡的存活和生长而言，慢速冷冻卵巢组织冷冻保存优于玻璃化冷冻。这些结果将有助于女性癌症患者的生育能力保存。
Two methods for the cryopreservation of human ovarian tissue were compared using a xenotransplantation model to establish a safe and effective cryopreservation method. Ovarian tissues were obtained from women who underwent benign ovarian surgery in the gynecology research unit of a university hospital. The tissues were transplanted into 112 ovariectomized female severe combined immunodeficient mice 4 weeks after slow freezing or vitrification cryopreservation. Tissues were retrieved 4 weeks later. Primordial follicular counts decreased after cryopreservation and xenotransplantation, and were significantly higher in the slow freezing group than in the vitrification group (p < 0.001). Immunohistochemistry and TUNEL assay showed that the Ki-67 and CD31 markers of follicular proliferation and angiogenesis were higher in the slow freezing group (p < 0.001 and p = 0.006, respectively) and DNA damage was greater in the vitrification group (p < 0.001). Western blotting showed that vitrification increased cellular apoptosis. Anti-Mullerian hormone expression was low in transplanted samples subjected to both cryopreservation techniques. Electron microscopy revealed primordial follicle deformation in the vitrification group. Slow freezing for ovarian tissue cryopreservation is superior to vitrification in terms of follicle survival and growth after xenotransplantation. These results will be useful for fertility preservation in female cancer patients.