title: Development of bovine embryos reconstructed by nuclear transfer of transfected and non-transfected adult fibroblast cells.
abstract: An association of two techniques, nuclear transfer (NT), and transfection of somatic animal cells, has numerous potential applications and considerable impact, mainly in agriculture, medicine, pharmacy, and fundamental biology. In addition, somatic cell nuclear transfer is the most efficient alternative to produce large transgenic animals. We compared in vitro and in vivo developmental capacities of NT using fibroblast cells isolated from a 14-month-old cloned Simmental heifer (FCE) vs the same line transfected with a plasmid containing neomycin-resistant genes (TFCE). There were no significant differences (P > 0.5) in either fusion (116/149 = 78% vs 216/301 = 72%), cleavage (78/116 = 67% vs 141/216 = 65%) and blastocyst (35/116 = 30% vs 52/216 = 24%) rates or in pregnancy rate at 30 to 35 days after embryo transfer (2/17 vs 3/17) between NT using FCE and TFCE, respectively. Transfection and long-term in vitro culture of transfected cells did not affect developmental capacity of NT embryos up to 40 days of gestation.
authors.affiliation: Embrapa Genetic Resources and Biotechnology, Parque Estação Biológica, Av. W5 Norte Final, 70770-900 Brasília, DF, Brazil.
authors.forename: Lilian T
authors.initials: LT
authors.lastname: Iguma
authors.forename: Sharon F C
authors.initials: SF
authors.lastname: Lisauskas
authors.forename: Eduardo O
authors.initials: EO
authors.lastname: Melo
authors.forename: Maurício M
authors.initials: MM
authors.lastname: Franco
authors.forename: Ivo
authors.initials: I
authors.lastname: Pivato
authors.forename: Giovanni R
authors.initials: GR
authors.lastname: Vianna
authors.forename: Regivaldo V
authors.initials: RV
authors.lastname: Sousa
authors.forename: Margot A N
authors.initials: MA
authors.lastname: Dode
authors.forename: Francisco J L
authors.initials: FJ
authors.lastname: Aragão
authors.forename: Elíbio L
authors.initials: EL
authors.lastname: Rech
authors.forename: Rodolfo
authors.initials: R
authors.lastname: Rumpf
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000502:physiology; D002417:Cattle / Q000235:genetics*; D002999:Clone Cells / Q000502:physiology; D019976:Cloning, Organism; D004624:Embryo Transfer; D047108:Embryonic Development / Q000502:physiology*; D005260:Female; D005347:Fibroblasts / Q000637:transplantation*; D053652:Nuclear Transfer Techniques; D016133:Polymerase Chain Reaction; D011247:Pregnancy; D014162:Transfection / Q000379:methods