人卵巢组织玻璃化冷冻方案的研究

发布时间：2018-12-29 16:21

【摘要】：目的:探讨适合人卵巢组织玻璃化冷冻的冷冻保护剂及合适载体。方法:收集40例因卵巢良性包块行手术者的卵巢皮质40片,将每片卵巢皮质切为20块卵巢组织块。将其随机分为10组,玻璃化冷冻组9组,另一组为对照组,9组玻璃化冷冻组再根据使用不同配伍浓度的冷冻保护剂(CPA1、CPA2、CPA3)及3种不同载体方法(微滴法、冻存管法、筛网法)进行细分。分析比较玻璃化冷冻组9个亚组与对照组冻融前后卵泡形态学变化、细胞增殖及凋亡程度。结果:1形态学HE染色示:仅CPA1组微滴法和筛网法、CPA2组微滴法与对照组比较,差异无统计学意义(P0.05)。2免疫组化示:仅CPA1组微滴法和筛网法卵巢组织卵泡的Ki-67表达与对照组比较,差异无统计学意义(P0.05)。3Tunel细胞凋亡检测示:各级卵泡中卵母细胞及颗粒细胞中均未见凋亡细胞。结论:低浓度冷冻保护剂DMSO联合EG(即CPA1液)使用微滴法或筛网为冷冻载体冻存卵巢组织,对卵巢组织的损伤最小。
[Abstract]:Objective: to study the suitable cryopreservation agent and carrier for vitrification of human ovarian tissue. Methods: 40 pieces of ovarian cortex were collected from 40 patients with benign ovarian mass. Each ovarian cortex was cut into 20 ovarian tissue blocks. They were randomly divided into 10 groups: vitrification group (n = 9) and control group (n = 9). According to the use of CPA1,CPA2,CPA3 and three different carrier methods (microdrop method, cryopreservation method), the vitrification group was treated with different concentrations of cryopreservation agent (CPA1,CPA2,CPA3). Screen method) for subdivision. The morphological changes, proliferation and apoptosis of follicles in 9 subgroups of vitrification group and control group were analyzed and compared before and after freezing and thawing. Results: 1Morphologic HE staining showed that only CPA1 group and sieve method were used, and CPA2 group was compared with control group. There was no significant difference (P0.05). 2 Immunohistochemistry showed that the expression of Ki-67 in ovarian follicles of CPA1 group was higher than that of control group. There was no significant difference (P0.05). Apoptosis of 3Tunel cells was not found in oocytes and granulosa cells in all follicles. Conclusion: cryopreservation of ovarian tissue with low concentration DMSO combined with EG (CPA1 solution) using microdrop method or sieve as cryopreservation carrier can cause the least damage to ovarian tissue.
【作者单位】：天津医科大学研究生院;南开大学;天津市中心妇产科医院;
【基金】：天津市卫生局资助项目(编号:12ZK101)
【分类号】：R714.8

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