| The study investigated somatic cell nuclear transfer in cashmere goat, including donor cell culture, recipient oocyte in vitro maturation, reconstructed oocyte electrofusion and activation, embryo in vitro culture and embryo transfer. The condition of 37℃,100% humidity and 5% CO2, DMEM/F12 supplemented with 10% FBS was used to support the goat skin fibroblasts growth in vitro. The method of explanted tissue culture can effectively establish the goat fibroblast cell line. The goat skin fibroblasts could be separated and purified by enzyme digestion. Growth curve analysis and chromosome counting can determine that the the isolated and cultured goat skin fibroblasts is similar with the growth characteristics of cultured cells in vitro (2n=60). The experiment also studied the collection methods of oocytes, the different serum. The result showed that there was significant difference between surface cutting and aspiration (P<0.05), EGS (natural estrus), EGS (PG-inducing estrus) and FBS (P<0.05). The conclusion was that surface cutting was more helpful for goat oocytes maturation; EGS (natural estrus) was significantly useful for oocytes maturation, homologous serum was better than heterologous serum for oocytes maturation. Matured goat oocytes were efficiently activated by Ionomycin together with 6-DMAP and 20.4% of blastocyst were achieved. It was higher than the rate of blastocyst achieved by 7% ethanol (3.2%) and electroactivation (9.3%). The goat parthenogenesis embryos were cultured 2～4h in 6-DMAP,we found that the blastocyst rate had no significant difference(P>0.05). When the reconstructed embryos were co-cultured with goat cumulus cell monolayer, the blastocyst development was 17.5%, but the blastocyst development was 15.4% without co-cultured, there is no significant difference. The fusion voltage 2400～3200V/cm, 30μs pulse duration, 2 pulse was suitable for the fusion of reconstructed embryos. A total of 140 reconstructed embryos were transferred into 11 recipients, no goat gestation.
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