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KMID : 1023620050290030175
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Reproductive and Developmental Biology
2005 Volume.29 No. 3 p.175 ~ p.180
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Effect of Demecolcine-Assisted Enucleation and Recipient Cell Cycle Stage on the Development of Nuclear Transfer Bovine Embryos
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Baek J. J.
Park C. K.
Yang Boo-Keun
Kim Choung-Ik
Cheong Hee-Tae
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Abstract
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This study was conducted to examine the effects of demecolcine-assisted enucleation and recipient cell cycle stage on the development of bovine somatic cell nuclear transfer (NT) embryos. In vitro cultured oocytes for h were classified by first polar body (1st PB) extrusion and cell cycle stage (MI and MII) and treated demecolcine for 40 min before enucleation. Enucleated oocytes were fused electrically with bovine ear skin cells, activated by Ca-ionophore+DMAP, and cultured in vitro. Most of eggs () treated with demecolcine protruded a chromosome mass and enucleated efficiently (, (P<0.05). Demecolcine did not have a deteriorative effect on the development of NT embryos. Developmental rate of NT embryos reconstituted with oocytes extruded 1st PB significantly higher than that of NT embryos produced by oocytes without 1st PB (, P<0.05). Cleavage and blastocyst formation rate of embryos reconstituted with MI oocytes (, respectively) were significantly lower than those of embryos reconstituted with MII oocytes (, respectively, P<0.05). From the present result, it is suggested that domecolcine is useful for the enucleation of recipient oocytes in bovine NT procedures, and MII oocytes rather than MI oocytes are more appropriate for recipient cytoplasm. Although, the potential to develop into blastocysts of NT embryos produced by 1st PB-nonextruded and MI oocytes was very low, these oorytes could be used for NT.
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KEYWORD
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Nuclear transfer, Enucleation, Demeco-lcine, Recipient cell cycle stage, In vitro development, bovine
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