title: Development of an optimized zona-free method of somatic cell nuclear transfer in the goat.
abstract: The purpose of this study was to develop an improved zona-free method of goat somatic cell nuclear transfer (SCNT) that has both ease of operation and efficiency. The main steps involved were: (1) optimization of in vitro oocyte maturation, (2) parthenogenetic activation of zona-free oocytes, (3) SCNT of zona-free anaphase II-telophase II (AII-TII) oocytes that subverted the need for long term UV-exposure of the oocytes, and (4) in vitro culture of groups of cloned embryos in wells in a highly efficient continuous serum-free embryo medium to the blastocyst stage before transfer to the recipients. Percentages of transgenic blastocyst production were 22.3 and 33.1% for adult and fetal cell lines, respectively. After transfer of cloned and transgenic blastocysts, 28.6 and 36.4% of the recipients were confirmed pregnant and 75 and 33.3% of the pregnancies resulted in the delivery of viable offspring, respectively. To our knowledge, this is the first report of successful live and survived birth of cloned and transgenic offspring through a whole procedure of in vitro oocyte maturation and embryo development to the blastocyst stage, and in this study the in vitro efficiencies of cloned and transgenic embryo production were higher than the available reports.
authors.affiliation: Department of Reproduction and Development, Cell Sciences Research Center, Royan Institute for Animal Biotechnology, ACECR, Esfahan, Iran. mh.nasr-esfahani@royaninstitute.org
authors.forename: M H
authors.initials: MH
authors.lastname: Nasr-Esfahani
authors.forename: S M
authors.initials: SM
authors.lastname: Hosseini
authors.forename: M
authors.initials: M
authors.lastname: Hajian
authors.forename: M
authors.initials: M
authors.lastname: Forouzanfar
authors.forename: S
authors.initials: S
authors.lastname: Ostadhosseini
authors.forename: P
authors.initials: P
authors.lastname: Abedi
authors.forename: Y
authors.initials: Y
authors.lastname: Khazaie
authors.forename: K
authors.initials: K
authors.lastname: Dormiani
authors.forename: K
authors.initials: K
authors.lastname: Ghaedi
authors.forename: M
authors.initials: M
authors.lastname: Forozanfar
authors.forename: H
authors.initials: H
authors.lastname: Gourabi
authors.forename: A H
authors.initials: AH
authors.lastname: Shahverdi
authors.forename: A D
authors.initials: AD
authors.lastname: Vosough
authors.forename: H
authors.initials: H
authors.lastname: Vojgani
mesh_terms: D000818:Animals; D030801:Animals, Genetically Modified; D001755:Blastocyst / Q000166:cytology* / Q000378:metabolism; D019976:Cloning, Organism; D005260:Female; D006041:Goats; D006801:Humans; D008297:Male; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology*; D010312:Parthenogenesis / Q000528:radiation effects; D011247:Pregnancy; D014466:Ultraviolet Rays