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Effect of Fusion Method and Passage Culture of Hanwoo (Korean Cattle) Ear Skin and Fetal Fibroblasts on the Development of Nuclear Transfer Embryos  

Yang Byoung-Chul (National Livestock Research Institute, RDA)
Im Gi-Sun (National Livestock Research Institute, RDA)
Lee Sang-Ki (National Livestock Research Institute, RDA)
Kim Se-Woong (National Livestock Research Institute, RDA)
Kim Dong-Hoon (National Livestock Research Institute, RDA)
Seong Hwan-Hoo (National Livestock Research Institute, RDA)
Yang Boh-Suk (National Livestock Research Institute, RDA)
Publication Information
Reproductive and Developmental Biology / v.30, no.1, 2006 , pp. 53-58 More about this Journal
Abstract
The study was conducted to evaluate the effects of culture period and fusion method on the development of somatic cell nuclear transfer (SCNT) embryos reconstituted with Korean bovine fetal fibroblast cells (KbFF) and Korean bovine adult ear skin fibroblast cells (KbESF). KbFF were isolated from a day 51 Korean cattle (Hanwoo) fetus, and KbESF were isolated from a 28 month old Hanwoo calf. The cells were cultured up to 15 weeks (passage 15) in vitro for SCNT. Chamber and electrode needles were used for comparing fusion of reconstituted eggs. The doubling times of KbFF and KbESF were 17.3 hr and 24.3 hr, respectively. The fusion and cleavage rates were significantly higher in needle group (76.1 and 81.2% respectively, P<0.05) than those in chamber group. However, the blastocyst development rate was not different between both groups. Fusion and cleavage rates of NT eggs reconstituted with KbESF did not affected by passage number, however, blastocyst rates were lower in passage $1{\sim}4$ group (21.3%) than passage $5{\sim}8$ (39.4%) and $13{\sim}15$ groups (40.4%, P<0.05). Whereas, fusion rate was lower in passage $1{\sim}4$ group (61.5%) than those of passage $5{\sim}8$(75.0%) and $13{\sim}15$ (76.8%) groups, but cleavage and blastocyst rates were similar regardless of passage number in the KbFF. The results suggest that fusion method can affect the development of SCNT embryos, whereas the long term culture up to 15 passages may not affect the development of SCNT embryos.
Keywords
Korean bovine ear skin fibroblast(KbESF); Korean bovine fetal fibroblast(KbFF); Nuclear transfer; Fusion; Passage number;
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