Cloning of pig embryo by using ear and tail fibroblasts as donor nuclei: Effect of time after maturation and pulse duration on the rate of fusion and development of embryo
2001
Parnpai, R. | Tasripoo, K. | Kamonpatana, M.
Present experiment was carried out to examine the time after maturation and different fusion parameters on the fusion and developmental rate of enucleated pig oocytes with donor cells. Donor cells were transferred into periviteline space of enucleated oocytes by micromanipulator. Fusion was performed by placing individual oocytes between electrodes to electrostimulate. In experiment 1, using two DC pulses at 20 V/oocyte either 10, 20, 30, 40 or 50 micro sec. The highest fusion rate was found when using pulse width at 50 micro sec in ear fibroblasts and tail muscle cells (71 and 61 percent). The highest rate of embryos developed to 8-cell stage was found when using pulse width at 50 micro sec in ear fibroblasts and tail muscle cells (21 and 18 percent). Embryos in this experiment could not develop beyond 8-cell stage. In experiment 2, fusion and developmental rate of embryos when using two DC pulses at 20 V/oocyte for 40 micro sec of oocytes cultured at 36, 41 and 46 h were not different among both types of donor cells. Our finding may be concluded that electrostimulation with two DC pulses at 20 V/oocyte for 40 and 50 micro sec was appropriated for fusion both types of donor cells with enucleated oocytes. Fusion and developmental rate of embryos when using two DC pulses at 20 V/oocyte for 40 micro sec of oocytes cultured at 36, 41 and 46 h were not different among both types of donor cells.
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