人卵巢组织冷冻保存及其评价方式的研究与进展

Cryopreservation of human ovarian tissue and the evaluation methods

背景:如何保存有生育需求的患者的卵巢功能使其能获得一定的生育能力,具有重要的意义及广阔的应用前景。目的:综合近年来卵巢组织冷冻的相关研究,对保存及评价方法作以综述,以探讨如何建立最佳的卵巢组织冷冻方案。方法:由第一作者检索1994年1月至2014年1月PubMed数据及万方数据库相关文献。英文检索词为ovarian tissue,ovarian tissue cryopreservation,freezing factors;中文检索词为卵巢组织,卵巢组织冷冻,冷冻影响因素。纳入与卵巢组织冷冻技术、影响冷冻化效果的因素、冷冻组织复苏及人卵巢组织移植相关的文献59篇进行分析。结果与结论:目前,卵巢组织冷冻主要分为慢速冷冻、快速冷冻及玻璃化冷冻技术,而不同的患者、组织切片的大小、冷冻保护剂种类、渗透时间及冷冻载体的不同,都会影响到冷冻效果。卵巢组织复苏后,主要通过组织学显微镜下观察卵泡及颗粒细胞的形态并进行计数,观察亚细胞结构的改变评价冻融效果;凋亡信号的检测;免疫组织化学分析检测复苏后增殖及凋亡相关的信息;体外培养检测内分泌水平的变化;基因水平的检测等。近年来,卵巢组织冷冻后的临床应用是将冷冻复苏后的组织进行移植,使患者恢复生殖内分泌功能,或获取成熟的生殖细胞,从而保存女性生育能力。然而,卵巢组织冷冻仍存在一些问题尚需进一步研究。

BACKGROUND：How to preserve the fertility and ovarian function in patients with procreation needs has important significance and broad application prospects. OBJECTIVE：To summarize the recent studies about ovarian tissue cryopreservation, preservation and evaluation methods, and to explore the best solution for ovarian tissue freezing. METHODS：An online computer-based retrieval of PubMed database and Wanfang database between January 1994 and January 2014 was performed by the first author. The key words were ovarian tissue, ovarian tissue cryopreservation, freezing factors in English and Chinese. Final y 59 literatures on the ovarian tissue cryopreservation technique, influencing factor of cryopreservation, resuscitation, and human ovarian tissue transplantation were included. RESULTS AND CONCLUSION：Currently, laboratory methods of ovarian tissue cryopreservation are divided into slow-rate freezing, ultra-rapid cooling and vitrification technology. The freezing effect may vary in different patients, tissue sections, cryoprotectants species, penetration time and frozen carrier. After the recovery, the histological morphology of granulosa cells and fol icles was observed. The fol icles were counted to evaluate ovarian tissue. Freeze-thaw effects were evaluated by observing the changes of subcellular structures. The apoptotic signals were also detected. Immunohistochemical analysis was applied to detect the proliferation after recovery and apoptosis-related information. Variation in the in vitro culture reflected endocrine levels. Genetic level detection was also performed. In recent years, frozen ovarian tissue is clinical y applied for the transplantation, to restore reproductive endocrine function of the patients, or get mature germ cells, thereby preserving female fertility. However, there are stil some questions about cryopreservation of ovarian tissue, which need further study.