两种冷冻保护剂玻璃化冷冻小鼠卵巢的比较研究

目的:探讨两种玻璃化冷冻保护剂对小鼠卵巢组织学和功能的影响。方法:将23只4周龄ICR雌鼠随机分为新鲜卵巢移植组(6只)、去势组(5只)、EG40组(6只)和ED20组(6只)。EG40组和ED20组分别应用乙二醇(EG)和联合应用EG与二甲基亚砜(DMSO)玻璃化冷冻小鼠卵巢组织,一周后解冻,将一部分复苏卵巢组织自体移植入小鼠肾被膜下,另一部分组织行组织学观察。移植术后5d开始观察所有小鼠的动情周期,一个月后处死小鼠,对存活的卵巢组织行组织学观察。结果:EG40组、ED20组和新鲜卵巢移植组小鼠动情周期出现率均为100%,出现动情周期的天数分别为10.2±1.2d、8.0±0.9d和6.8±1.0d,EG40组动情周期出现天数明显多于新鲜卵巢移植组(P<0.01);ED20组与新鲜卵巢移植组差异无显著性(P>0.05)。移植存活的卵巢组织内可见不同发育阶段的卵泡,形态正常,但冻融卵巢组织内卵泡数量比新鲜组织少。结论:联合应用玻璃化冷冻保护剂EG和DMSO对小鼠卵巢组织学和功能的影响较小。

Effects of Two Cryoprotectants on Mouse Ovarian Tissue after Vitrification

Objective: To observe the effects of two cryoprotectants on morphology and function of mouse ovarian tissues after vitrification. Methods: A total of 23 four-week old ICR female mice were randomly assigned to four groups: non-frozen (n=6), ovariectomized (n=5), EG40 (n=6) and ED20 (n=6). Ovarian tissues were vitrified with ethylene glycol (EG) or EG combined with dimethyl sulphoxide (DMSO) as cryoprotectants. Some of the vitrified tissues were transplanted under kidney capsules, the others were examined histologically after frozen and thawed. Estrus cycle was observed 5 days after ovarian autografting. The recovered autografts were collected and examined histologically one month after transplantation. Results: In the EG40, ED20 and non-frozen groups, the initiation rate of estrus were all 100%, but the initiation days of estrus were 10.2±1.2 d, 8.0±0.9 d and 6.8±1.0 d respectively. The initiation days of estrus in the EG40 group were significantly more than that in the non-frozen group (P<0.01). There was no significant difference between the ED20 and the non-frozen groups (P>0.05). The follicles at different development stages in autografts were normal morphology, but the number of follicles in vitrified ovarian tissues was less than that in non-frozen tissues. Conclusion: The vitrification with EG and DMSO as cryoprotectants could effectively preserve mouse ovarian tissues.