玻璃化冷冻保存对小鼠卵巢组织的影响

Effects of Cryopreservation on Mouse Ovarian Tissue after Vitrification

目的探讨玻璃化冷冻保存对小鼠卵巢组织内卵泡形态及卵巢组织激素分泌功能的影响。方法将23只4周龄ICR雌鼠随机分为新鲜卵巢组、去势组和玻璃化冷冻组,应用乙二醇(EG)和二甲基亚砜(DMSO)玻璃化冷冻小鼠卵巢组织。冷冻前后取部分小鼠卵巢组织行组织学观察;同时将部分新鲜和复苏组织自体移植入小鼠肾被膜下。移植术后5 d开始观察小鼠的动情周期,术后30d测定小鼠血清雌激素浓度。结果冻融卵巢组织内存活的原始卵泡和初级卵泡与新鲜组织无显著差异(P>0.05);而次级卵泡的存活率显著下降(P<0.01)。移植术后玻璃化冷冻组和新鲜卵巢组小鼠均出现动情周期,两组动情周期出现时间无明显差异(P>0.05)。移植术后第30天两组小鼠血清雌激素浓度无显著差异(P>0.05)。结论玻璃化冷冻法可以较好地保存小鼠卵巢组织内的原始卵泡和初级卵泡,且不影响卵巢组织的激素分泌。

Objective To observe the effects of cryoprezervation on morphology of various stage follicles and estradiul secretion of mouse ovarian tissues after vitrification. Methods 23 four-week old ICR female mice were randomly assigned to three groups： non-frozen （n = 8）, ovariectomized （n = 5） and vitrified （n = 10）. Ovarian tissues were vitrified with ethylene glycol （EG） combined with dimethyl sulphoxide （DMSO） as cryopretectants. Some of the non-frozen and frozen tissues were examined histologically, the others were transplanted under mice kidney capsules. Estrus cycles were began to observe 5 days after ovarian transplantation, and mouse serum estradiol were detected 30 days after transplantation. Results The number of primordial follicles and that of primary follicles in vitrified tissues were not significant different from that in non-frozen tissues respectively. However, the percent viability of secondary follicles were （71.76 ± 7.90） % in frozen tissues, compared with （91.67 ± 6.36） % in non-frozen tissues （ P 〈 0.01 ）. In the vitrified and non-frozen groups, the initiation rate of estrus were all 100% , and the initiation days of estrus were （7.6 ± 1.1 ）days and （6.6 ± 0.9） days respectively （P 〉 0.05 ） . There was no significant difference in estradiol production of ovarian tissues 30 days after transplantation between the frozen and non-frozen groups. Conclusion Vitrified mouse ovarian tissue could maintain similar viability of primordial follicles and primary follicles and estradiol secretion with non-frozen ovarian tissue.