Somatic Nuclear Transplantation and Serial Nuclear Transplantation of Human Finger-domain Lacking t-PA Gene in Goat

To research developmental competence of transgenic somatic cell by serial nuclear transplantation, goat cloned embryos were compared with recloned embryos in ability of in vitro development. Fetal fibroblasts including human finger-domain lacking t-PA gene was microinjected into the cytoplasm of the MII oocytes. Goat embryos (G0) were cloned by this procedure. A single blastomere from 16-cell to 64-cell goat cloned embryos (G0) was microinjected into the Intracytoplasm of the MII oocytes. Goat embryos (G1) were cloned by this procedure. Goat embryos (G2, G3) were recloned using 16-cell to 64-cell recloned embryos. The developmental time of donor embryo affected the developmental rate of recloned embryos (G1, G2). The result indicates: the cleavage rate of cloned embryos (G0) (76.45±1.17%) was no difference significantly with recloned embryos (G1 G2 G3) (72.18%±1.97%, 76.05%±2.38%, 75.99%±2.84%). The developmental rate of morulae and blastocysts of cloned embryos (47.20%±2.93%, 11.00%±1.42%) were higher than of recloned embryos (34.99%±2.66%, 28.23%±2.00%, 23.34%±1.99%) (3.87%±0.67%, 2.08%±1.66%, 0); The morulae rate (29.57%±1.53%, 24.43%±1.87%), blastocysts rate (1.96%±1.31%, 2.01%±1.34%) of recloned embryos (G1 G2) from 16-cell recloned embryos was lower than that (34.32%±1.31%, 29.76%±1.66%), (3.86%±1.03%, 3.48%±0.34%) from 32-64-cell recloned embryos (P>0.05). In conclusion, nuclear transfer embryos were not recloned mostly; Embryos recloned using 32-cell to 64-cell embryos achieved higher developmental ability compared with using 16-cell embryos.