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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。
It is hoped that the production of transgenic tree shrew animal model can be achieved through the method of mature culture of tree oocytes and in vitro fertilization. Using gonadotropin to supervise tree shrews, TCM199 completely cultured the oocytes matured in vitro. Oocytes and in vitro fertilized epididymal sperm were fertilized in vitro, and fertilized eggs developed into morula and blastula. The results showed that the oocyte maturation rate of A grade 76.7%, B grade 55.01%, C grade 17.39%. Fertilization rate of 52%, somatic cell co-culture of fertilized eggs and non-co-culture of the cleavage rates were 37.14% and 9.6%. The morula / blastocyst rates were 13.57% and 0, respectively (P <0.05). Tree 鼩 oocyte maturation, in vitro fertilization experimental method feasible, fertilized eggs in vitro can develop to the blastocyst stage.