J Vet Sci.  2011 Mar;12(1):83-89. 10.4142/jvs.2011.12.1.83.

Cloned calves derived from somatic cell nuclear transfer embryos cultured in chemically defined medium or modified synthetic oviduct fluid

• Jang G
• Hong SG
• Lee BC

• KMID: 1067341
• DOI: http://doi.org/10.4142/jvs.2011.12.1.83

Abstract

Somatic cell nuclear transfer (SCNT) is considered to be a critical tool for propagating valuable animals. To determine the productivity calves resulting from embryos derived with different culture media, enucleated oocytes matured in vitro were reconstructed with fetal fibroblasts, fused, and activated. The cloned embryos were cultured in modified synthetic oviduct fluid (mSOF) or a chemically defined medium (CDM) and developmental competence was monitored. After 7 days of culturing, the blastocysts were transferred into the uterine horn of estrus-synchronized recipients. SCNT embryos that were cultured in mSOF or CDM developed to the blastocysts stages at similar rates (26.6% vs. 22.5%, respectively). A total of 67 preimplantational stage embryos were transferred into 34 recipients and six cloned calves were born by caesarean section, or assisted or natural delivery. Survival of transferred blastocysts to live cloned calves in the mSOF and the CDM was 18.5% (to recipients), 9.6% (to blastocysts) and 42.9% (to recipients), 20.0% (to blastocysts), respectively. DNA analysis showed that all cloned calves were genetically identical to the donor cells. These results demonstrate that SCNT embryos cultured in CDM showed higher viability as judged by survival of the calves that came to term compared to blastocysts derived from mSOF cultures.

Keyword

bovine embryo; cloned calves; culture media; implantation; somatic cell nuclear transfer

MeSH Terms

Animals
Blastocyst/physiology
*Cattle
Cloning, Organism/methods/*veterinary
Culture Media/chemistry
Embryo Culture Techniques
Embryo Transfer
Embryonic Development
Female
Fertilization in Vitro/*veterinary
Nuclear Transfer Techniques/*veterinary
Pregnancy

Figure

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  Fig. 1 Cloned calves resulting from somatic cell nuclear transfer. Two were born on June 8, 2006 by natural delivery (A) and assisted delivery (B). A cloned calf (C) was born on June 10, 2006 by natural delivery. A cloned calf was delivered by caesarean section on May 3, 2006 (D). Twin cloned calves were born by assisted delivery (F). One of twins died from severe diarrhea and the other lived (E). Another set of twin cloned calves (G) were born prematurely at 264 days after embryo transfer by assisted delivery but they died from respiratory distress.

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