SOME FACTORS AFFECTING IN VITRO DEVELOPMENT OF PORCINE EMBRYO RECONSTITUTION FROM SOMATIC CELLS NUCLEAR TRANSFER
ZHANG De Fu WANG Ying CHEN Yin WANG Kai Karl Schellander LIN Cai Lu ( Amnnal Husbondry and Veterrnary Research Institulc, Shanghar Academy Academy of Agrtcultral Sciences, Shanghat 201106; Division of Anintal Gertelre Engineermg, Shanghar Mwtierpal Key Laboratory,of Agr-Genetrts and Breeding, Shanghar 201106; Shanghai Municipal Key Laboratory of Deoelopmental Bology, Shanghai No 2 Medical University, Shanghai 2110025: Anial Breeding Institute, Bomn University, Bomn 53121. Gerinany)
In this paper, a study on reconslitution 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 to morula( 11.7%) and blastocysls(6.7% ) phases which were higher than those derived from the fusion with FC( p0. 05) . The results of this study also involved the effects of oocyte collection method and maturational age of recipient oocytcs during the in vitro develpoment of nuclear-transfer embryos which were reconstructed with cuhured cumulus cells. The cumulus cells sync hronized in G0/G1 phases through serum-starvation culture, were transferred into enucleated oocytes which were collected by aspiration or dissection method and culinred 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 oocytes collection methods, activation treatment in the presence of cytochalasm H 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 was significantly higher( p0. 05) by activation with the combination of electric pulsation and 6-DMAP. These results suggest that reconstiluted porcine embryos derived from cultured cumulus cells can develop to the blastocyst stage and that the development of the former could be improved by reconstruction with young oocyte cytoplasl after the activation with the combination of electric pulsation and 6-DMAP.