云岭山羊卵母细胞玻璃化冷冻的研究

摘要

试验以屠宰场云岭黑山羊卵巢卵母细胞为材料,研究其玻璃化冷冻的效果.试验中选用20％ EG+20％ DMSO为冷冻液、冷冻环为载体,以20 s、40 s玻璃化时间冷冻GV和MⅡ期的卵母细胞.结果表明,GV期卵母细胞的形态正常率、成熟率和卵裂率都很低,且解冻成熟培养后冷冻组的成熟率和卵裂率极显著低于对照组(P＜0.01).而MⅡ期卵母细胞冷冻效果较好,毒性试验组和冷冻组形态正常率分别为91.1％和83.3％,明显高于GV期；孤雌激活后毒性组卵裂率与对照组无显著性差异(P＞0.05),冷冻组的卵裂率显著低于对照组(P＜0.05).用20 s、40 s玻璃化时间冷冻的卵母细胞解冻后GV和MⅡ期各组均无显著差异.根据试验结果得出在冷冻保存中最好冷冻MⅡ期的卵母细胞,以便提高后期的卵裂率和囊胚率；卵母细胞玻璃化时间在40 s内均不影响卵母细胞的活力和发育潜力.%The experimental material used Yunling goat oocytes from slaughterhouse,to study the effect of the oocytes vitrification. Using 20% EG+20% DMSO as a frozen solution and the cryloop for carrier to vitrify the GV and MⅡ stage oocytes in 20 s and 40 s vitrification time. The results showed that,GV oocytes morphologically normal rate,maturation rate and cleavage rate were very low. After maturation culture,the maturation rate and cleavage rate of the vitrification groups was significantly lower than the control group (P<0. 01). MⅡ oocytes frozen was better, the rate of normal morphology of the toxicity test group and vitrification group were 91. 1 % and 83. 3% , was significantly higher than the GV stage. After parthenogenetic activation,the cleavage rate,the toxicity test group and control group,were no significant differences(P>0. 05). The vitrification group cleavage was significantly lower than the control group(P<0. 05). The GV and M Q oocytes in each group were not significantly different in 20 s and 40 s vitrification time. According to the results it was the best to vitrify M Ⅱ oocytes in the cryopreservation.in order to improve the cleavage rate and blastocyst rate. The oocytes vitrification time did not affect oocyte viability and developmental potential in the 40 s inside.