title: Cloned transgenic calves produced from nonquiescent fetal fibroblasts.
abstract: An efficient system for genetic modification and large-scale cloning of cattle is of importance for agriculture, biotechnology, and human medicine. Here, actively dividing fetal fibroblasts were genetically modified with a marker gene, a clonal line was selected, and the cells were fused to enucleated mature oocytes. Out of 28 embryos transferred to 11 recipient cows, three healthy, identical, transgenic calves were generated. Furthermore, the life-span of near senescent fibroblasts could be extended by nuclear transfer, as indicated by population doublings in fibroblast lines derived from a 40-day-old fetal clone. With the ability to extend the life-span of these primary cultured cells, this system would be useful for inducing complex genetic modifications in cattle.
authors.affiliation: Department of Veterinary and Animal Sciences, University of Massachusetts, Amherst, MA 01003, USA.
authors.forename: J B
authors.initials: JB
authors.lastname: Cibelli
authors.forename: S L
authors.initials: SL
authors.lastname: Stice
authors.forename: P J
authors.initials: PJ
authors.lastname: Golueke
authors.forename: J J
authors.initials: JJ
authors.lastname: Kane
authors.forename: J
authors.initials: J
authors.lastname: Jerry
authors.forename: C
authors.initials: C
authors.lastname: Blackwell
authors.forename: F A
authors.initials: FA
authors.lastname: Ponce de León
authors.forename: J M
authors.initials: JM
authors.lastname: Robl
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst; D002417:Cattle / Q000196:embryology / Q000235:genetics*; D002455:Cell Division; D002467:Cell Nucleus / Q000235:genetics; D002478:Cells, Cultured; D016922:Cellular Senescence; D002999:Clone Cells; D019976:Cloning, Organism; D004624:Embryo Transfer; D005260:Female; D005333:Fetus / Q000166:cytology; D005347:Fibroblasts / Q000166:cytology*; D016193:G1 Phase; D008297:Male; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology; D014162:Transfection; D019076:Transgenes