title: A comparison of two in vitro maturation media for use with adult porcine oocytes for adult somatic cell nuclear transfer.
abstract: Two media used to mature adult porcine oocytes for somatic cell nuclear transfer were compared. In the first experiment, parthenogenetic embryos were produced using a maturation medium used by us previously to clone pigs (OMM199) and that described by Kühholzer et al. (2001) to transport oocytes overnight (BOMED). There was no difference in maturation rates between the two different media. However, BOMED medium increased the percentage of parthenogenetic embryos that developed to the blastocyst stage compared with OMM199 (49% vs. 29%, respectively). In a second experiment, BOMED medium increased the percentage of SCNT embryos that developed to the blastocyst stage compared with OMM199 (22% vs. 8%, respectively). The efficiency of our cloning protocol using adult oocytes matured in BOMED medium was then determined by transferring SCNT embryos reconstructed using adult fibroblasts to synchronized recipients. Primary cultures of adult fibroblasts were obtained from two adult male pigs and used for SCNT (passages 2-4). Between 82 and 146 fused couplets were transferred to seven recipients synchronized 1 day behind the embryos. Five recipients (71% pregnancy rate) subsequently farrowed a total of 23 piglets (4.4 average litter size). Overall efficiencies (liveborn/embryos transferred) were 3.2% for all transfers and 4.3% for animals that gave birth.
authors.affiliation: Reproductive Biotechnology Group, Research Centre for Reproductive Health, Discipline of Obstetrics and Gynaecology, School of Paediatrics and Reproductive Health, University of Adelaide, Adelaide, South Australia, Australia. luke.beebe@adelaide.edu.au
authors.forename: Luke
authors.initials: L
authors.lastname: Beebe
authors.forename: Stephen
authors.initials: S
authors.lastname: McIlfatrick
authors.forename: Christopher
authors.initials: C
authors.lastname: Grupen
authors.forename: Andrew
authors.initials: A
authors.lastname: Boquest
authors.forename: Sharon
authors.initials: S
authors.lastname: Harrison
authors.forename: Renate
authors.initials: R
authors.lastname: Faast
authors.forename: Rodney
authors.initials: R
authors.lastname: Ashman
authors.forename: James
authors.initials: J
authors.lastname: Wengle
authors.forename: Hamish
authors.initials: H
authors.lastname: Hamilton
authors.forename: Mark
authors.initials: M
authors.lastname: Nottle
mesh_terms: D000818:Animals; D002467:Cell Nucleus / Q000378:metabolism; D019976:Cloning, Organism / Q000379:methods*; D003470:Culture Media / Q000494:pharmacology*; D004624:Embryo Transfer / Q000379:methods; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D005821:Genetic Techniques; D066298:In Vitro Techniques; D053652:Nuclear Transfer Techniques; D009865:Oocytes / Q000166:cytology* / Q000378:metabolism; D011247:Pregnancy; D011270:Pregnancy, Animal; D013552:Swine
chemical_list: D003470:Culture Media