树鼩卵母细胞的体外成熟培养及体外受精

利用树鼩卵母细胞的成熟培养、体外受精等方法,生产树鼩的试管婴儿,期望通过此路径来实现生产转基因树鼩动物模型。使用促性腺激素进行树鼩超数排卵、TCM199完全培基对卵母细胞进行体外成熟培养、卵母细胞与体外获能的附睾精子进行体外受精、受精卵发育至桑椹胚、囊胚的实验。结果表明,卵母细胞成熟率A级76.7%、B级55.01%、C级17.39%。受精率52%,受精卵采用体细胞共培养与非共培养的分裂率分别为37.14%和9.6%。桑椹胚/囊胚发育率分别为13.57%和0,(P<0.05)。树鼩卵母细胞的成熟培养、体外受精实验方法可行,受精卵在体外能发育到囊胚阶段。

Study on in vitro maturation and fertilization of Tupaia glis oocytes

In order to produce transgenetic Tupaia glis animal model successfull y, the methods to produce IVF Tupaia glis, such as IVM and IVF, has been explore d. Superovulation, IVM with TCM199, IVF between oocytes and in vitro capacitati on sperms from epididymis has been investigated.Oocytes maturation rate were A g rade76.7%and B grade 55.01% and C grade 17.39%,respectively; fertilization rate was 52%. Cleavage rate of the zygotes co-cultured with somatic cells and zygotes non-co-cultured were 18.5% and 9.6%, respectively; morula and blastocyst rate w ere 8.6 and 1.2, repectively. ( p<0.05) . Considering the zygote can develop to blastocyst in vitro , the IVM and IVF of Tupaia glis oocytes is feasible. The r esults of the experiment can provide effective foundation to transgenetic Tupaia glis animal model production.