摘要
目的探讨应用自制冷冻环进行人卵巢组织玻璃化冷冻的效果。方法2008年5月至9月在华中科技大学同济医学院附属同济医院收集5例人卵巢组织标本,以乙二醇、二甲基亚砜、蔗糖作为冷冻保护剂,自制冷冻环为载体进行玻璃化冷冻。并观察卵巢新鲜组织和冻融后组织始基卵泡和初级卵泡的形态学变化、凋亡情况、超微结构变化及体外培养内分泌功能。结果新鲜及冻融后组织形态正常卵泡比值分别是89.46±4.94、84.47±4.66,无统计学意义(P>0.05)。凋亡面积分别是(0.07±0.02)%、(0.10±0.05)%,无明显增加(P>0.05)。超微结构无明显改变。新鲜组织及冻融后组织体外培养上清液雌二醇含量分别(2549.73±711.87)pmol/L、(2514.87±714.66)pmol/L,无统计学意义(P>0.05)。结论自制冷冻环可作为一种便宜、方便、可行的人卵巢组织低温保存的可替代方法。
Objective To investigate cryopreservation effects of human ovarian tissue using self-made cryoloop. Methods Ovarian biopsies collected from 5 women of Tongji Hospital Affiliated to Tongji Medical College of Huazhong University of Science and Technology from May to September in 2008 were vitrificated by cryoprotectants,composed of ethylene glycol,dimethylsuphoxide and sucrose,using self-made cryoloop as a cooling device. The morphology analysis of primordial and primary follicles,apoptosis signals,ultastructure,endocriological function cultured in vitro of the fresh and vitrificated/thawed ovarian tissues were accessed. Results The ratio of morphologically normal primordial and primary follicle（84.47±4.66）was not different compared with the fresh one（89.46±4.94）,P0.05. The mean TUNEL-positive surface area values of the fresh tissue and the vitrificated one were（0.07±0.02）%,（0.10±0.05）%,and no statistically significant difference was found（P0.05）. The ultrastructure was similarly preserved after vitrification. After culture of fresh tissue pieces and vitrificated/thawed pieces,the supernatants showed average estrodiol concentration of （2549.73±711.87）pmol/L,（2514.87±714.66）pmol/L,respectively,and no statistically significant difference was found（P0.05）. Conclusion Cryoloop vitrification is an effective,simple and inexpensive alternative for human ovarian tissue cryopreservation.
出处
《中国实用妇科与产科杂志》
CAS
CSCD
北大核心
2010年第1期39-42,共4页
Chinese Journal of Practical Gynecology and Obstetrics
基金
湖北省科技攻关课题(2007AA301B38-1)
关键词
卵巢组织
人
玻璃化冷冻
卵泡
体外培养
超微结构
ovarian tissue
human
vitrification
cryoloop
follicle
in vitro culture
ultrastructure
