Development of interspecies cloned monkey embryos reconstructed with bovine enucleated oocytes.
This study was carried out to determine whether culture media reconstructed with bovine enucleated oocytes and the expression pattern of Oct-4 could support dedifferentiaton of monkey fibroblasts in interspecies cloned monkey embryos. In this study, monkey and bovine skin fibroblasts were used as donor cells for reconstruction with bovine enucleated oocytes. The reconstructed monkey interspecies somatic cell nuclear transfer (iSCNT) embryos were then cultured under six different culture conditions with modifications of the embryo culture media and normal bovine and monkey specifications. The Oct-4 expression patterns of the embryos were examined at the two-cell to blastocyst stages using immunocytochemistry. The monkey iSCNT embryos showed similar cleavage rates to those of bovine SCNT and bovine parthenogenetic activation (PA). However, the monkey iSCNT embryos were not able to develop beyond the 16-cell stage under any of the culture conditions. In monkey and bovine SCNT embryos, Oct-4 could be detected from the two-cell to blastocyst stage, and in bovine PA embryos, Oct-4 was detectable from the morula to blastocyst stage. These results suggested that bovine ooplasm could support dedifferentiation of monkey somatic cell nuclei but could not support embryo development to either the compact morula or blastocyst stage. In conclusion, we found that the culture conditions that tend to enhance monkey iSCNT embryo development and the expression pattern of Oct-4 in cloned embryos (monkey iSCNT and bovine SCNT) are different than in bovine PA embryos.
Lorthongpanich
Chanchao
C
Embryo Technology and Stem Cell Research Center, Suranaree University of Technology, Nakhon Ratchasima, Thailand.
Laowtammathron
Chuti
C
Chan
Anthony Wing Sang
AW
Ketudat-Cairns
Mariena
M
Parnpai
Rangsun
R
D000818:Animals; D002417:Cattle; D002459:Cell Fusion; D002467:Cell Nucleus / Q000187:drug effects; D002678:Chimera / Q000196:embryology; D019976:Cloning, Organism / Q000379:methods; D003470:Culture Media / Q000494:pharmacology; D046149:Embryo Culture Techniques; D004622:Embryo, Mammalian / Q000166:cytology / Q000187:drug effects / Q000378:metabolism / Q000502:physiology; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology*; D008252:Macaca fascicularis / Q000196:embryology*; D008954:Models, Biological; D053652:Nuclear Transfer Techniques; D050814:Octamer Transcription Factor-3 / Q000378:metabolism; D009865:Oocytes / Q000166:cytology*
D003470:Culture Media; D050814:Octamer Transcription Factor-3