不同玻璃化冷冻载体对人卵巢组织冷冻效果的影响

摘要

Objective:To compare the effects of different vitrification carriers on the freezing human ovarian tissue,and provide the theoretical and experimental basis in selecting frozen solution. Methods:The human ovarian cortex tissues were cut into the size of 10 mmí1 mmí1 mm,randomly divided into the fresh group(group A),needle immersed vitrification group(group B),solid surface vitrification group( group C) and direct covered vitrification group( group D) according to different vitrification carriers. The fresh group were fixed by 10% formaldehyde,embeded in paraffin,sectioned and stained with hematoxylin and eosin. The other three groups were preserved in liquid nitrogen after freezing for two months, then fixed, sectioned and stained using rapid rewarming protocol. The proportions of normal primordium follicles in group B,C and D were calculated by histological analysis,which was used to analyze the frozen effects. Results:There were 809 primordial follicles in four groups,the proportions of normal primordium follicles in group A,B, C and D were 96. 12%,88. 14%,80. 00% and 81. 25%,respectively. The proportions of normal primordium follicles in group B,C and D were lower than that in group A(P0. 05). Conclusions:Compared with the fresh tissue,the proportions of normal primordium follicles decrease after freezing,but the vitrification freezing can be used to preserve the human ovarian tissue still.%目的：：比较不同的玻璃化冷冻载体对人卵巢组织冷冻效果的影响,为人卵巢组织玻璃化冷冻方案的选择提供理论基础和实验参考依据。方法：预处理人卵巢皮质切成10 mm×1 mm×1 mm大小,随机分为新鲜组( A组)、针浸润玻璃化冷冻组(B组)、表面玻璃化冷冻组(C组)和直接覆盖玻璃化冷冻组(D组)。 A组直接予以固定、组织切片及HE染色；B、C、D 3组采用不同玻璃化冷冻载体冷冻后,投入液氮保存2个月,采用快速复温法解冻后予以固定、切片及染色。行形态学分析,通过计算各组卵巢组织切片中正常形态始基卵泡率,比较各组冷冻效果。结果：4组共计数809个始基卵泡,其中正常形态始基卵泡率分别为：A组96.12%,B组88.14%,C 组80.00%,D组81.25%。行玻璃化冷冻的3组卵巢组织正常形态始基卵泡率均明显小于A组(P0.05)。结论：玻璃化冷冻后人卵巢组织正常形态始基卵泡率较新鲜组织下降,但仍能较好地保存人类卵巢组织,可用于人类卵巢组织的冷冻保存。