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KMID : 0900920000240010059
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Korean journal of Animal Reproduction
2000 Volume.24 No. 1 p.59 ~ p.67
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Nuclear Remodeling and In Vitro Development of Bovine Oocytes Following Nuclear Transfer of Bovine Fetal Fibroblasts
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Um J.-H.
Uhm S.-J.
Kim N.-H.
Lee H.-T.
Chung K.-S.
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Abstract
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This study was investigated the developmental potential of bovine embryos following nuclear transfer with bovine fetal fibroblasts (BFF). BFF were isolated from a male 45-day-old-fetus. Non-starved BFF labeled with MitoTracker were transferred into perivitelline space of enucleated oocytes. BFF-oocyte units were fused by electric pulse, and then fused oocytes were activated with calcium ionophore A23187 and subsequently 6-dimethylaminopurine (6-DMAP). The resulting zygotes were placed into CRlaa bovine embryo culture medium. Transfer of the nucleus into enucleated oocyte led to premature chromosome condensation, swelling and pronucleus formation. Remodeled oocytes were developed to the mitotic and 2-cell stage at 18 to 26 h after nuclear transfer. The incidence of in vitro development to the blastocyst stages was 21% of fused oocytes. Mitochondria of BFF eliminated rapidly and were not detected at 8 h after fusion. These results suggest that BFF can be successfully reprogrammed in enucleated bovine oocytes, and that reconstructed embryos can develop to the blastocyst stage.
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KEYWORD
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Bovine, Development, Mitochondria, Nuclear transfer, Remodeling
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