以细胞筛为载体的玻璃化冷冻和慢速程序化冷冻保存人卵巢组织的效果比较

发布时间：2018-02-28 12:23

本文关键词： 细胞筛玻璃化冷冻慢速冷冻卵巢冷冻生育力保存　出处：《生殖医学杂志》2016年09期 　论文类型：期刊论文

【摘要】：目的比较以细胞筛为载体的玻璃化冷冻与慢速程序化冷冻用于人卵巢组织冷冻的效果。方法人卵巢组织取皮质切块后,随机分为3组:新鲜组织对照组(F组)、慢速程序化冷冻组(S组)及以细胞筛为载体玻璃化冷冻组(V组)。卵巢组织冷冻复苏后,固定切片后行苏木素-伊红染色,观察卵泡形态;使用TdT介导的dUTP缺口末端标记技术,观察卵泡凋亡情况;部分卵巢组织体外培养,隔日采集培养液后检测雌二醇(E2)浓度。比较三组卵泡正常形态率、卵泡凋亡率及E2浓度。结果与F组原始卵泡正常形态率(91.1%)相比,V组原始卵泡正常形态率(88.1%)无显著差异(P0.05),而S组原始卵泡正常形态率(79.6%)显著下降(P0.001);V组初级卵泡正常形态率(74.2%)与S组(73.6%)相似,但两组均低于F组(89.0%)(P0.05);凋亡检测中,3组凋亡率无显著差异(P0.05);体外培养2周,各组E2浓度持续上升,F组E2浓度显著高于S组、V组(P0.001),而S组与V组E2浓度无显著差异(P0.05)。结论以细胞筛为载体的玻璃化冷冻能较好地保存人卵巢组织,复苏后组织体外培养后,还可持续分泌E2。
[Abstract]:Objective to compare the effects of vitrified cryopreservation and slow programmed cryopreservation on human ovarian tissue. They were randomly divided into three groups: fresh tissue control group (group F), slow programmed freezing group (group S) and vitrified cryopreservation group (group V) with cell sieve as carrier. Ovarian tissue was frozen and resuscitated, fixed sections were stained with hematoxylin and eosin, and follicular morphology was observed. TdT mediated dUTP Nick end labeling technique was used to observe follicular apoptosis, partial ovarian tissue was cultured in vitro and estradiol E2 concentration was measured after the next day culture medium. The normal morphological rate of follicles was compared among the three groups. Results there was no significant difference in normal morphology rate of primordial follicles (P 0.05) and normal morphology rate of primordial follicles in group S (79.6%) compared with group F (91.1%). Form rate (74.2) and group S (73.6). But both groups were lower than F group (P 0.05), the apoptosis rate of the three groups was not significantly different from that of F group (P 0.05), and cultured in vitro for 2 weeks. The concentration of E2 in group F was significantly higher than that in group S (P 0.001), but there was no significant difference between group S and group V (P 0.05). Conclusion the vitrified cryopreservation with cell sieve as carrier can better preserve human ovarian tissue, and after resuscitation, the tissue can be cultured in vitro after resuscitation. E2 can also be secreted sustainably.
【作者单位】：中国医学科学院北京协和医学院北京协和医院妇产科;
【基金】：2015年度默克雪兰诺中国生殖医学研究基金
【分类号】：R714.8

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