水牛卵母细胞玻璃化冷冻保存  

Vitrificated cryopreservation of in vitro matured buffalo oocytes

作　　者：卞桂华[1] 冯贵雪[1] 陈玉[1] 杨素芳[1] 王晓丽[1] 石德顺[1] 

机构地区：[1]广西大学动物繁殖研究所,广西南宁530005

出　　处：《中国兽医学报》2009年第1期110-112,120,共4页Chinese Journal of Veterinary Science

基　　金：广西青年基金项目(桂科青0542022);国家863资助项目(2002AA206651)

摘　　要：以水牛MII期的卵母细胞为材料,利用玻璃化冷冻液EDS33(16.5%EG+16.5%DMSO+sucrose)对水牛MII期的卵母细胞进行两步法(玻璃毛细管(GMP)和拉细的开口塑料细管(OPS))玻璃化冷冻保存,即卵母细胞首先放入预平衡液(7.5%EG+7.5%DMSO+sucrose)中平衡3 min,再移入玻璃化冷冻液中30 s后装管直接投入液氮。解冻是在蔗糖浓度逐渐降低的解冻液中进行的。解冻后存活的卵母细胞孤雌激活,通过囊胚发育率作为评定卵母细胞冷冻效果的指标。结果发现,GMP和OPS法冷冻保存的水牛卵母细胞解冻后的存活率(分别为96.80%和97.41%)与对照组卵母细胞的存活率(100%)3者之间差异均不显著(P>0.05)。GMP法和OPS法冷冻的水牛卵母细胞激活后的胚胎分裂率和囊胚发育率2者均明显低于对照组(分别为30.58%和28.32%vs50.94%,10.81%和9.38%vs 29.63%,P<0.05),而这2种方法冷冻的水牛卵母细胞激活后的分裂率和囊胚发育率差异均不显著(P>0.05)。这表明GMP和OPS玻璃化冷冻方法可以用于水牛卵母细胞的冷冻,并且玻璃化冷冻的卵母细胞能继续分裂并发育到囊胚。The present study was conducted to evaluate the feasibility of cryopreserving in vitro matured buffalo oocytes by the glass micropipette （GMP） and open pulled straw （OPS）. The buffalo oocytes were cryopreserved with EDS33（16.5% ethylene glycol and 16.5% dimethyl sulfoxide and sucrose）in GMP and OPS. In the two-step method,oocytes were first pretreated with 7.5% EG and 7.5% DMSO for 3 minutes, then vitrified in 16.5% EG and 16.5 % DMSO for 30s and warmed in decreasing concentrations of sucrose. The oocytes that survived for vitrification were parthenogenetically activated and cultured in vitro up to the blastocyst stage. Results indicated that survival rate of buffalo oocytes did not differ significantly in GMP,OPS and the control groups （96.80% ,97.41% vs. 100%,P〉0.05）. However, oocytes vitrified by the GMP and OPS methods had a significantly lower cleavage and blastocyst rates than the control （30.58 % and 28.32 % vs. 50.94 %;10.81% and 9. 38% vs. 29.63 %, respectively; P〈0.05）, while cleavage and blastocyst rates of the surviving vitrified oocytes （parthenogenetically activated） were approximative in GMP and OPS groups （P〉0.05）.

关 键 词：GMP OPS 玻璃化 冷冻保存 卵母细胞 水牛 

分 类 号：S823.83[农业科学—畜牧学]