摘要
目的:优化性成熟SD大鼠超排卵效果及建立大鼠成熟卵母细胞冷冻方法.方法:对8周龄SD大鼠腹腔注射15 IU/100 g孕马血清促性腺激素(PMSG),48 h后注射2种不同剂量人绒毛膜促性腺激素(HCG),15 IU/100 g及7.5 IU/100 g进行超排,20~22 h后输卵管收集卵母细胞卵丘复合体,脱颗粒细胞获得形态良好的成熟卵子,比较不同剂量HCG的超排卵母细胞数,并通过玻璃化冷冻法对卵母细胞进行冷冻,根据不同冷冻时间分为5组,观察冷冻解冻后卵子存活率.结果:7.5 IU/100 g HCG剂量平均超排42.2枚卵母细胞,显著高于15 IU/100 g HCG剂量组;冷冻9 min组(94.29%)及冷冻6 min组(94.37%)玻璃化冷冻卵母细胞存活率最高,显著高于冷冻15 min组(50.70%)和冷冻3 min组(48.57%),显著高于冷冻12 min组(68.06%).结论:性成熟大鼠在15 IU/100 g PMSG及7.5 IU/100 g HCG剂量组合下,可获得最佳超排效果,玻璃化冷冻9 min组及冷冻6 min组能更好地保存大鼠卵母细胞.
Objective:To optimize the effect of superovulation in adult SD rats and establish a method for vitrifying mature oocytes in rats.Methods:Eight-week-old SD rats were intraperitoneally injected with 15 IU/100 g of pregnant mare serum gonadotropin(PMSG).After 48 h,two different doses of human chorionic gonadotropin(HCG)(15 IU/100 g and 7.5 IU/100 g)were injected for superovulation.After 20-22 h,the oocyte cumulus complex was collected from the fallopian tube,and the granulosa cells were denuded to obtain mature oocytes with good morphology.The numbers of superovulated oocytes of different doses of HCG were compared.The oocytes were frozen by vitrification,and divided into five groups according to different timing.The survival rates of oocytes after vitrification were observed.Results:An average of 42.2 oocytes were collected in the 7.5 IU/100 g HCG dose group,which was significantly higher than that of 15 IU/100 g HCG.The highest survival rates of oocytes were in the 9 min vitrifi cation group(94.29%)and 6 min vitrification group(94.37%),which were significantly higher than the 15 min vitrification group(50.70%)and 3 min vitrification group(48.57%),higher than 12 min vitrification group(68.06%).Conclusion:Adult rats could obtain optimal superovulation effect under the combination of 15 IU/100 g PMSG and 7.5 IU/100 g HCG dose.9 min vitrification group and 6 min vitrification group can better preserve the rat oocytes which provides a guarantee for the subsequent scientific research of rat oocytes.
出处
《解剖学杂志》
CAS
2019年第6期543-546,共4页
Chinese Journal of Anatomy
基金
上海中医药大学2016年预算内项目（2016N47）
作者简介
尹萍,E-mail:120042559@qq.com;通信作者:童国庆,E-mail:drivftongguoqing@hotmail.com。
