Study On Boer Goat Somatic Cell Nuclear Transfer

The study investigated somatic cell nuclear transfer in Boer goat,including donorcell culture, the β-galactosidase expression with fibroblast in vitro culture, recipientoocyte in vitro maturation, and used somatic cell from adult goat as donor to cloneBoer Goat. The method of tissue block culture and enzyme digestion can effectivelyestablish the goat fibroblast cell line.Growth curve analysis can determined that theisolated and cultured Boer goat skin fibroblasts is similar with the growthcharacteristics of cultured cells in vitro. Cell that be thawed can maintain the samegrowth rate with that before be frozen, it indicated that the condition of freezing andthe procedure of thawing are appropriate to this experiment.The fibroblasts in vitroculture were proliferated and passaged till50generation, and there were not appearobviously groups proliferation repose in50generation., it showed that the subculturecan continue in theory. Age have an important effect on the aging of fibroblasts invitro cultured, The positive rate of older group was higher than young group,andyoung group was higher than juvenile group at the same generation (P<0.01). Thenumber of cell generation increased, the number of senescent cell increased, and thereis extremely significant correlation between the cell generation and the number ofsenescent cell(P<0.01). The positive rate between10and15generation of juvenilegoat have significant difference(P<0.05),the positive rate between15and20generations of youth ewes had not difference(P>0.05). At any ages, from20generations the positive rate between adjacent two generation have extremelysignificant difference (P<0.01), it showed that the ordinal number of passage havesignificant effect on fibroblasts aging. Gender have significant effect on fibroblastaging at10to45generation, positive rate of rams group were lower than ewes groupat the same generation(P<0.01), but they didn't have difference at50generation(P>0.05). Both slaughterhouse ovaries and ovarium that be primed by FSHsuperovulation, the maturation rate of oocytes in vitro cultured of grade A and Boocyte were higher than grade C(84.1%,77.4%vs44.8%;70.3%,65.5%vs41.7%),and they had extremely significant differences(P<0.01), it indicated that the grade of oocyte has significant effect on maturation rate of oocytes in vitro cultured, but therewere not differences between grade A and B (P>0.05). The percentage of grade A andB oocyte in the total oocytes that have been obtained from FSH superovulation andslaughterhouse were86.2%and58.3%, every ovarium can be obtained9.6and4.1oocytes on average, respectively,grade A and B oocyte were8.3and2.4.,it indicatethat the oocytes from FSH superovulation were more than that fromslaughterhouse(P<0.05), these showed that FSH superovulation can distinct increasethe oocytes quantity and quality. The maturation rate of oocytes in vitro cultured fromFSH superovulation was higher than that from slaughterhouse (81.6%vs67.6%), andthere were extremely significant difference(P<0.01), it indicated that FSHsuperovulation treatment can improve the maturation rate of oocytes in vitro culturedand oocytes developmental capacity. Finally, we used somatic cell from adult goatthat had been cultured for a long period as donor and cloned Boer Goat successfully,then established a complete nuclear transfer procedure.