不同方法冻融山羊卵巢组织的研究

目的探讨山羊卵巢组织不同玻璃化冷冻方法对其卵母细胞体外成熟、体外受精及胚胎发育的影响。方法采用三种不同的冷冻保护液(Ⅰ、Ⅱ、Ⅲ)保存的卵巢组织吸入0.25ml麦管中直接投入液氮中冷冻保存。采用慢速解冻法解冻后对卵巢组织进行形态学观察;收集卵母细胞进行体外培养。结果三种不同冷冻方法冷冻卵巢组织解冻后收集的卵母细胞存活率分别为55.2%、48.9%和46.7%,组间比较无显著性差异(P>0.05);成熟率分别为58.5%(I组)、28.9%(II组)、33.3%(III组),I组显著高于Ⅱ、Ⅲ组(P<0.01);三组的受精率、卵裂率、8细胞胚胎的形成率均无显著性差异(P>0.05);卵巢组织解冻后形态学分析表明,I组效果好于Ⅱ、Ⅲ组。结论采用玻璃化方法冷冻卵巢组织,在冷冻保护液中加入卵黄可提高冷冻保护效果。

Study on different protocols for goat ovarian tissue cryopreservation

Objective:The present study was conducted to investigate the effects of cryopreservation protocols of goat ovarian tissues on in vitro maturation(IVM),in vitro fertilization(IVF) of oocytes and embryos development.Methods:The ovarian tissues were vitrified with three different cryoprotectants(I,II,III).Cryopreservation of ovarian pieces was accomplished by plunging 0.25ml straws into liquid nitrogen.Histological observation of ovarian tissues was performed after thawing,and the oocytes recovered from these tissues were cultured to observe the maturation and fertilization of the oocytes and embryo development.Results:The survival rates of the oocytes recovered from the tissues vitrified with cryoprotectant I,II and III were 55.2%,48.9% and 46.7%,respectively.There was no significant difference among them(P>0.05).The IVM rate in Group I was 58.5%,significantly higher than that of Group II(28.9%) and Group III(33.3%)(P<0.01).There was no significant difference in the rates of IVF,cleavage and 8-cell between groups(P>0.05).Histological observation showed that the tissues in Group I were better cryopreserved than those in Group II and GroupIII.Conclusion:The ovarian tissues could be effectively vitrified with the cryoprotectant containing egg yolk.