Vitrification and xenografting of human ovarian tissue
Vitrification and xenografting of human ovarian tissue
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DOI：
10.1016/j.fertnstert.2012.07.1109
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发表时间：
2012-11-01
影响因子：
6.7
通讯作者：
Van Langendonckt, Anne
中科院分区：
文献类型：
--
作者：
Amorim, Christiani Andrade;Dolmans, Marie-Madeleine;Van Langendonckt, Anne
目的：评估两种玻璃化冷冻方案在异种移植模型中冷冻保存人腔前卵泡的效率。设计：初步研究。设置：大学医院的妇科研究单位。患者：从7名年龄在30-41岁的女性中获得卵巢活检。干预：卵巢组织碎片进行三种冷冻保存方案之一（缓慢冷冻、玻璃化冷冻方案1和玻璃化冷冻方案2）并异种移植至裸鼠1周。通过组织学分析确定冷冻保存和移植后形态正常的卵泡数量和纤维化表面积。TUNEL法检测细胞凋亡。还进行了TUNEL阳性表面积的形态学分析。结果：异种移植后，观察到不同冻存程序之间的差异。根据TUNEL分析，两种玻璃化冷冻方案均显示出比常规冷冻方法更好的腔前卵泡保存。此外，组织学评价显示，玻璃化冷冻（方案2）-温热的卵巢组织中原始卵泡的比例显著高于冷冻-解冻的组织。生长卵泡和纤维化表面积的比例是相似的，在所有groups. Conclusions（S）：玻璃化冷冻程序出现保存不仅是形态和生存的腔前卵泡后1周的异种移植，但也有能力恢复folliculogenesis。此外，玻璃化冷冻方案2对异种移植后原始卵泡的静止状态有积极的影响。（Fertil Steril（R）2012;98：1291-8. (C)美国生殖医学会（American Society for Reproductive Medicine）
Objective: To assess the efficiency of two vitrification protocols to cryopreserve human preantral follicles with the use of a xenografting model.Design: Pilot study.Setting: Gynecology research unit in a university hospital.Patient(s): Ovarian biopsies were obtained from seven women aged 30-41 years.Intervention(s): Ovarian tissue fragments were subjected to one of three cryopreservation protocols (slow freezing, vitrification protocol 1, and vitrification protocol 2) and xenografted for 1 week to nude mice.Main Outcome Measure(s): The number of morphologically normal follicles after cryopreservation and grafting and fibrotic surface area were determined by histologic analysis. Apoptosis was assessed by the TUNEL method. Morphometric analysis of TUNEL-positive surface area also was performed. Follicle proliferation was evaluated by immunohistochemistry.Result(s): After xenografting, a difference was observed between the cryopreservation procedures applied. According to TUNEL analysis, both vitrification protocols showed better preservation of preantral follicles than the conventional freezing method. Moreover, histologic evaluation showed a significantly higher proportion of primordial follicles in vitrified (protocol 2)-warmed ovarian tissue than in frozen-thawed tissue. The proportion of growing follicles and fibrotic surface area was similar in all groups.Conclusion(s): Vitrification procedures appeared to preserve not only the morphology and survival of preantral follicles after 1 week of xenografting, but also their ability to resume folliculogenesis. In addition, vitrification protocol 2 had a positive impact on the quiescent state of primordial follicles after xenografting. (Fertil Steril (R) 2012;98: 1291-8. (C) 2012 by American Society for Reproductive Medicine.)