供体细胞来源及预处理对牛体细胞核移植效率的影响

以牛外耳皮肤成纤维细胞为核供体细胞,比较了不同同步化诱导方式(血清饥饿法与接触抑制法)、供体细胞冷冻与否、供体动物性别与年龄等因素对体细胞核移植效率的影响。结果表明,接触抑制法处理的供体细胞核移植胚囊胚率显著高于饥饿法(P<0.05);未冷冻的供体细胞核移植胚囊胚率显著高于冷冻供体细胞(P<0.05);成年母牛供体的囊胚率显著高于成年公牛(P<0.05);不同年龄供体细胞的核移植胚囊胚率差异不显著(P>0.05)。试验表明,以接触抑制法诱导未冷冻成年母牛外耳皮肤成纤维细胞为核供体,有利于核移植胚囊胚的发育。

Effects of derivation and pretreatment of donor cells on the efficiency of nuclear transfer in cattle

In this study,with bovine auricle fibroblasts used as nuclear donor,the effects of nuclear donor with serum starvation vs.contact inhibition,frozen vs.non-frozen,as well as the age and sex of animals from which nuclear donor cells were derived on the efficiency of nuclear transfer were investigated and compared.The results showed that the rates of blastocysts derived from nuclear donor cells pretreated with contact inhibition or non-freezing were significantly higher than those derived from nuclear donor cells pretreated with serum starvation or frozen respectively(P<0.05).Moreover,when female cells were used as nuclear donors,the blastocyst rate of cloned embryos was significantly higher than those derived from male cells(P<0.05).However,the age of animal from which donor cells derived could not affect the efficiency of nuclear transfer(P>0.05).When adult female bovine auricle fibroblasts were pretreated with contact inhibition and were not frozen,the results indicated those were significantly beneficial to the development of nuclear transfer embryos.