不同冷冻保护剂组合及平衡时间对玻璃化保存卵巢组织的影响.DOC

不同冷冻保护剂组合及平衡时间对玻璃化保存卵巢组织的影响 山东省临沂市妇幼保健院生殖中心 王新民 276001 [摘要] 目的：比较不同冷冻保护剂组合及平衡时间对玻璃化保存卵巢组织的影响[Ⅰ:20% (v/v) 乙二醇 + 20% (v/v) 二甲基亚砜,Ⅱ: 20% (v/v) 乙二醇 + 20% (v/v) 丙二醇, Ⅲ:20% (v/v) 丙二醇 + 20% (v/v) 二甲基亚砜] 和不同的平衡时间 ( a: 15分钟, b: 30 分钟, c： 45 分钟) 将小鼠随机分成不同的实验组。结果：在所有的实验组中，Ⅱb组的始基卵泡和窦前卵泡正常形态率最高，卵泡凋亡率和Bax蛋白表达率最低。结论：玻璃化冷冻保护液为 [20% (v/v) 乙二醇 + 20% (v/v) 丙二醇]及平衡时间为30分钟的方案是玻璃化冻存小鼠卵巢组织的适宜方案。 [关键词]：冷冻保护剂；组合；平衡时间；玻璃化；卵巢组织 中图分类号：R332 文献标识码：A Effects of different combinations of cryoprotectants with different equilibrium time on the mouse ovarian tissue during vitrification WANG Xin-min Center of Reproduction Medicine, Linyi Maternal and Child Health Hospital of Shang Dong province, 276001,China. E-mail:xuefr190@yeah,net [Abstract] Objective:The aim of this study is to detect the effects of different combinations of cryoprotectants with different equilibrium time on the mouse ovarian tissue during vitrification. Methods:Ovarian tissue of mice was vitrified-thawed. Mice (n=80) were randomly assigned to treatment groups according to different vitrification solution [Ⅰ:20% (v/v) EG + 20% (v/v) DMSO,Ⅱ: 20% (v/v) EG + 20% (v/v) PROH, Ⅲ:20% (v/v) PROH + 20% (v/v) DMSO] and different length of equilibrium time ( a: 15minutes, b :30 minutes, c 45 minutes). Results: The rates of morphologically normal primordial follicle and preantral follicle in GroupⅡb were the highest in all these treatment groups. The rates of apoptotic follicle and follicle with Bax protein expression in GroupⅡb were the lowest in all these treatment groups. Conclusion: vitrification solution [20% (v/v) EG + 20% (v/v) PROH] with equilibrium time of 30 minutes is optimal selection for vitrifying mouse ovarian tissue. Keywords: cryoprotectant; Combination; equilibrium time; vitrification; ovarian tissue 随着医学水平的迅速发展,恶性肿瘤患者的长期生存率得到明显提高,但是大剂量的放化疗对患者的内分泌功能和生育能力造成严重影响。卵巢组织冷冻保存是一种较理想的卵巢功能保存方法[1]。目前常用的冷冻方法是程序化冷冻，但它需要昂贵仪器，费时，消耗液氮多[2]。而玻璃化冷冻是一种简单、高效、快速的冷冻方法。在超低温条件下，高浓度的玻璃化冷冻液渗入到细胞内部，使之处于玻璃化状态而不形成冰晶，因此对细胞的损伤较小[3]。1985年Rall等首先使用玻璃