【摘要】 以卵丘细胞为核供体细胞组成重构胚 ,卵裂率达到 5 6.7% ,发育至桑椹胚率达到1 1 .7% ,囊胚率为 6.7% ,显著高于成纤维细胞重构胚 (P <0 .0 5 )。本文还研究了卵母细胞的采集方法、激活程序和卵龄对卵丘细胞核移植重构胚体外发育的影响。以血清饥饿法将卵丘细胞诱导至G0 G1 期 ,抽吸法 解剖法采集卵母细胞 ,体外培养 3 3～ 44h ,将卵丘细胞放至去核卵母细胞的卵周隙中 ,重构胚以钙离子载体A2 3 81 7或电脉冲结合 6 DMAP激活处理 ,体外培养 6d。研究表明 ,卵母细胞采集方法、激活液中细胞松弛素 (CB)、激活程序并不影响重构胚的发育 (以卵龄 44h的卵母细胞为受体 ) ;而以电脉冲结合 6 DMAP激活处理能提高重构胚发育能力 (以卵龄 3 3h的卵母细胞为受体 ) (P <0 .0 5 )。本研究显示 ,以电脉冲结合 6 DMAP激活卵丘细胞重构胚 ,体外能发育至囊胚更多还原

【Abstract】 In present paper,a study on reconstitution of porcine oocytes by using nuclear transfer with cumulus cells(CC) and fibroblast cells(FC) was carried out.Reconstituted oocytes which were the fusion with CC and showed a cleavage rate of 56.7%,developed into morula (11.7%) and blastocysts (6.7%) phases which were higher than those derived from the fusion with FC( P <0.05).The results of this study also involved the effects of oocyte collection method,activation protocol and maturational age of recipient oocytes during the in vitro develpoment of nuclear transfer embryos which were reconstituted with cultured cumulus cells.The cumulus cells synchronized in G 0/G 1 phases through serum starvation culture,were transferred into enuclated oocytes which were collected by aspiration or dissection method and cultured for 33 or 44 h.Reconstituted embryos were activated with a combination of calcium ionophore A23187 or electric pulsation and 6 DMAP,and cultured for 6 days.As for the oocyte collection methods,activation treatment in the presence of cytochalasin B and activation protocols did not affect the developmental rate of embryos reconstituted with 44 h mature recipients.However,the development rate of reconstituted embryos with 33 h mature recipients were significantly higher( P <0.05) by activation with the combination of electric pulsation and 6 DMAP.These results suggest that the reconstituted porcine embryos derived from cultured cumulus cells can develop into the blastocyst stage and that the development of the former could be improved for the reconstitution with young oocyte cytoplast after the activation with the combination of electric pulsation and 6 DMAP.更多还原