清洗后石蜡油对昆明小鼠胚胎培养及其发育的影响

Effect of washing mineral oil on development of mouse embryos in vitro and in vivo after embryo transfer

目的建立有效而简便的石蜡油清洗回收系统,并检验其对昆明白小鼠胚胎培养的效果。方法石蜡油分为:对照组、回收组、医用组及化学纯组。除对照组之外各组均行清洗石蜡油三步处理回收法。手术收集小鼠1-细胞阶段的胚胎,用TE培养液培养,观察胚胎发育情况并进行胚胎移植。结果石蜡油回收组培养,从2-细胞到囊胚的比例均高于对照组,囊胚的形成速度比对照组快。囊胚率回收组为23%,对照组17%。回收组胚胎移植的产仔率为44%、对照组为16%;清洗与未清洗医用石蜡油都能支持胚胎发育到囊胚;化学纯轻质石蜡油经清洗后可有效提高囊胚的发育率。并提高胚胎及胎盘的重量。结论本石蜡油清洗回收系统效果良好,适用于胚胎培养。

Objective： To establish a simple and effective washing procedure for both new and used mineral oil. Methods： In our protocol, oil was first mixed with 95% ethanol at 1 ： 1 for 5 min. The recovered oil was then heated to boil with 0.31 mol/L NaCl for 30 rain and separated. Finally, the separated oil was treated with anhydrous sodium sulfate overnight and filtered for use. The KM strain mouse 1-cell embryos were collected surgically and cultured in droplets of medium overlaid with washed or unwashed mineral oil of different types. Sigma mineral oil M8410 was used as control. Cleavage rate and blastocyst formation were recorded daily and some embryos were transferred into recipients. Results： It was shown that washed recycled oil can promote the embryo development from 2-cell to blastocyst stage （23.3%） when compared with that of control （16.9 %）. Offspring were obtained at 44% （7/16） and 16 % （3/19） of transferred embryos from washed recycled oil and control respectively. Both washed and unwashed mineral oil of medical grade can support KM mouse embryo development from 1-cell to blastocyst stage. Washing treatment to chemical grade light mineral oil can increase the development rate of embryos at each stage and the weight of fetus and placenta. Conclusion： The washed mineral oil by our washing procedure is beneficial for improving the embryo development.