不同采集方法对猪卵母细胞的采集效率和成熟率的影响

The Collection Efficiency and Maturation Rate of Porcine Oocytes Influenced by the Collection Methods

本研究旨在开发出一种对COCs损伤小且采集效率和卵母细胞成熟率高的卵母细胞采集方法。从屠宰场采集来的卵巢运回实验室后随机分为3组,分别用抽吸法、解剖法和刀切过滤法3种方法采集COCs,然后进行体外成熟培养,通过对COCs采集效率、COCs完整性及卵母细胞体外成熟率的比较,研究解剖法、刀切过滤法和抽吸法3种采集方法对猪卵母细胞的采集效率和成熟率的影响。结果表明:每个卵巢采集COCs个数,刀切过滤法显著高于解剖法和抽吸法(P<0.05),而解剖法仅为1.55个,显著低于抽吸法(P<0.05);采集效率抽吸法最佳,采集和挑选单个COCs的时间显著低于解剖法和刀切过滤法(P<0.05),刀切过滤法次之,但显著低于解剖法(P<0.05);A、B级COCs比例,解剖法高达99.23%,显著高于其他2种方法(P<0.05),刀切过滤法与抽吸法差异不显著(P>0.05);卵母细胞的成熟率,解剖法和刀切过滤法都达到80%,二者间差异不显著(P>0.05),但显著高于抽吸法(P<0.05);100个成熟卵母细胞所需的卵巢数量,刀切过滤法最少,仅为17.4个,而解剖法和抽吸法分别高达80.6和90.3个;需要的总采集时间(采集和挑选的时间),刀切过滤法最低,仅为126.6min,而解剖法最高,为758.9min。综上所述,与抽吸法和解剖法相比,刀切过滤法是一种高效、快速的卵母细胞采集方法。

In order to improve the effect of oocyte collection method,we investigated the collection efficiency,the proportion of A and B grade oocytes and in vitro maturation rate of pig oocytes collected by the dissection method,the knife cutting filtration method and the aspiration method.The number of pig oocytes obtained from each ovary by knife cutting filtration method was significantly higher than those by the dissection method and the aspiration method;the time of collecting and selecting ovary（collection efficiency） by the aspiration method was less than those by the knife cutting filtration method and the dissection method;the proportion of A and B grade oocytes by the dissection methods,reached to 99.23%,was significantly higher than those by the knife cutting filtration method and aspiration method;the maturation rate of pig oocytes by the dissection method and the knife cutting filtration method,reached to 80%,was higher than that by the aspiration method;to obtain 100 oocytes,the least needed ovaries by the knife cutting filtration method was only 17.4 compared with 80.6 and 90.3 by the dissection method and aspiration method.It took only 126.6 min for the whole collection by the knife cutting filtration method,however 758.9 min by the dissection method.The results indicate that the knife cutting filtration method is an efficiency and rapid method for the collection of porcine oocytes in comparison with the aspiration method and the dissection method.