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Production of Cloned Calves by the Transfer of Somatic Cells Derived from Frozen Tissues Using Simple Portable $CO_2$ Incubator

  • Dong, Y.J. (Faculty of Animal Science, Raiyo Agricultural College) ;
  • Bai, X.J. (Faculty of Animal Science, Raiyo Agricultural College) ;
  • Varisanga, M.D. (Faculty of Science, Technology and Environmental Studies, The Open University of Tanzania, Dar es Salaam) ;
  • Mtango, N.R. (The United Graduate School of Veterinary Sciences, Yamaguchi University, Laboratory of Animal Reproduction & Applied Biotechnology) ;
  • Otoi, T. (The United Graduate School of Veterinary Sciences, Yamaguchi University, Laboratory of Animal Reproduction & Applied Biotechnology) ;
  • Rajamahendran, R. (Departments of Animal Science, The University of British Columbia) ;
  • Suzuki, T. (The United Graduate School of Veterinary Sciences, Yamaguchi University, Laboratory of Animal Reproduction & Applied Biotechnology)
  • Received : 2003.02.03
  • Accepted : 2003.10.11
  • Published : 2004.02.01

Abstract

The ability of frozen-thawed fetal skin was examined to generate viable cell lines for nuclear transfer. Fetal skin frozen at -20$-20^{\circ}C$, $-30^{\circ}C$ or $-80^{\circ}C$ in the presence of 5% DMSO used as tissue explants to generate somatic cells. The resultant confluent cells were then used as donors for nuclear transfer (NT). Of the bovine NT embryos reconstracted from the somatic cells, 62.3%, 76.6% to 65% showed cleavage 70.5%, 81.9% to 78.5% reached the stage of morula formation and 39.7%, 43.2% or 47.6% reached the blastocyst stage. There was no significant difference in development when the NT embryos were compared with those reconstracted from fresh somatic cell derieved skin tissues (72%, 75.3%, and 45.2%, for cleavage, and development to morula and blastocyst stage, respectively). NT embryos were then placed in a portable $CO_2$ incubator and carried to China from Japan by air. After reaching to farm, two NT embryos were transferred to each of 5 recipients. We obtained 2 NT calves which birth weights is 30kg and 36kg female, and gestation periods is 281 and 284 days, respectively. There were no observation any abnormality from those calves. The results indicated that cell lines derieved from bovine fetal skin cryopreserved by a simple method could be used as donors in nuclear transfer using the portable $CO_2$ incubator.

Keywords

Cloning;Frozen Fetal Skin;Electrofusion;Portable ${CO}_2$ Incubator;Blastocysts;Calves

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