Experimental Research on the Construction of Cloned Embryos Through Human-rabbit Inter-species Nuclear Transfer

Therapeutic cloning is one of our major research objectives, and therapeutic cloning is based on human somatic cell nuclear transfer. Though inter-species nuclear transfer has been introduced to construct human somatic cell cloned embryos, the effects of type, passage and preparation method of donor cells on embryo development remain to be determined. In our experiment, cloned embryos were reconstructed with different passage and preparation methods of ossocartilaginous cell, skin fibroblast and cumulus cells. The cumulus cell embryos show significantly higher development rates than the other two (P<0.05). The development rate of embryos reconstructed with skin fibroblasts of different passage number and somatic cells of different chilling durations show no significant difference. Also, FISH were conducted to detect nuclear derivation of the embryos. The result showed that the nuclei of the inter-species cloned embryo cells came from human. We conclude (1) that cloned embryos can be constructed through human-rabbit inter-species nuclear transfer; (2) different kinds of somatic cell result in different efficiency of nuclear transfer, while in vitro passage of the donor does not influence embryo development; (3) refrigeration is a convenient and efficient donor cell preparation method. Finally, it is feasible to detect DNA genotype through FISH.