小鼠卵巢组织玻璃化冷冻保存的研究

为了探索小鼠卵巢组织玻璃化冷冻保存的有效方法,以5周龄昆明小白鼠的卵巢为材料,分别采用不同的冷冻保护剂、平衡时间、冷冻载体和组织块大小对小鼠卵巢组织进行玻璃化冷冻保存。玻璃化冷冻保存的基本程序:取小白鼠卵巢,切块,玻璃化冷冻。玻璃化冷冻效果评判的方法是解冻玻璃化冷冻保存的卵巢组织块,收集其中的未成熟卵,体外培养和体外受精,观察其体外成熟、体外受精及所形成胚胎的发育能力。结果表明,小鼠卵巢组织玻璃化冷冻保存以卵巢组织块大小为1mm3、以半麦管作为载体、在75mL/LED(EG+DMSO)中平衡15min、以用150mL/L ED玻璃化冷冻液处理3min的方法,冷冻保存效果最好。

Cryopreservation of mouse ovarian tissue by vitrification

To establish an effective cryopreservation method for mouse ovarian tissue,the ovarian tissues from five-week-old Kunming mice were cryopreserved by using different cryoprotectants,equilibrium time,carrier and the size of the ovarian tissue.Basic procedure of ovarian tissue cryopreservation was as follows:cutting the ovarian into tissue pieces,putting into carrier and cultivated in vitro cryopreserving by vitrification.The methods of evaluating results of cryopreservation were as follows:thawing the cryopreserved mouse ovarian tissue,isolating immature oocytes from mouse ovarian tissue,culturing the immature oocytes to maturation in vitro,fertilizing mature oocytes,in vitro,cultivating fertilized oocytes in vitro,and observing in vitro maturation and fertilization ability of isolating oocytes and developmental competence of the fertilized oocytes.The results showed that the optimum procedure cryopreservation of ovarian tissue by vitrification was:cutting ovary into 1 mm3,pretreating with 75 mL/L EG for 15 min,treating with 150 mL/L ED EG for 3 min,and using hemi-straw as carrier.