title: A novel embryo culture media supplement that improves pregnancy rates in mice.
abstract: The preimplantation embryo in vivo is exposed to numerous growth factors in the female reproductive tract, which are not recapitulated in embryo culture media in vitro The IGF2 and plasminogen activator systems facilitate blastocyst development. We hypothesized that the addition of IGF2 in combination with urokinase plasminogen activator (uPA) and plasminogen could improve rates of blastocyst hatching and implantation in mice. B6BcF1 and CBAB6F2 mouse embryos were divided into one of four supplemented culture media treatment groups: (1) control (media only); (2) 12.5 nM IGF2; (3) 10 µg/mL uPA and 5 µg/mL plasminogen; or (4) a combination of IGF2, uPA and plasminogen treatments. Embryo development to blastocyst stage and hatching were assessed before transfer to pseudopregnant recipient females and implantation, pregnancy rates and postnatal growth were assessed. After 90.5 h of culture, IGF2 + U + P treatment increased the percentage of B6BcF1 embryos that were hatching/hatched and percentage developing to blastocyst stage compared with controls (P < 0.02). Following B6BcF1 embryo transfer, IGF2 + U + P treatment increased implantation sites at day 8 of pregnancy compared with controls (P < 0.05). Replication in the CBAB6F2 mouse strain showed significant improvements in pregnancy rates at days 8 and 18 but not in blastocyst development. No adverse effects were seen on gestational age, litter size or birthweight, or the reproductive capacity of offspring of IGF2 + U + P treated embryos. For embryos susceptible to detrimental effects of in vitro culture, IGF2, uPA and plasminogen supplementation of culture media can improve pregnancy success, but the effect of treatment is dependent on the mouse strain.
authors.affiliation: Adelaide Medical School.
authors.forename: A R
authors.initials: AR
authors.lastname: Highet
authors.affiliation: School of AgricultureFood and Wine, Robinson Research Institute, The University of Adelaide, Adelaide, South Australia, Australia.
authors.forename: T
authors.initials: T
authors.lastname: Bianco-Miotto
authors.affiliation: Adelaide Medical School.|Priority Research Centre for Reproductive ScienceSchool of Biomedical Sciences and Pharmacy, University of Newcastle, Newcastle, New South Wales, Australia.
authors.forename: K G
authors.initials: KG
authors.lastname: Pringle
authors.affiliation: Adelaide Medical School.
authors.forename: A
authors.initials: A
authors.lastname: Peura
authors.affiliation: Bioinformatics FacilityRobinson Research Institute, The University of Adelaide, Adelaide, South Australia, Australia.|Institute for Molecular BioscienceUniversity of Queensland, St Lucia, Queensland, Australia.
authors.forename: S
authors.initials: S
authors.lastname: Bent
authors.affiliation: Adelaide Medical School.
authors.forename: J
authors.initials: J
authors.lastname: Zhang
authors.affiliation: Adelaide Medical School.
authors.forename: M B
authors.initials: MB
authors.lastname: Nottle
authors.affiliation: Adelaide Medical School.|Australian Research Council Centre for Excellence in Nanoscale BiophotonicsAdelaide, South Australia, Australia.
authors.forename: J G
authors.initials: JG
authors.lastname: Thompson
authors.affiliation: Adelaide Medical School claire.roberts@adelaide.edu.au.
authors.forename: C T
authors.initials: CT
authors.lastname: Roberts
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000166:cytology* / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology*; D046149:Embryo Culture Techniques; D010064:Embryo Implantation / Q000187:drug effects; D004624:Embryo Transfer; D047108:Embryonic Development / Q000187:drug effects; D005260:Female; D005307:Fertilization in Vitro / Q000187:drug effects*; D007335:Insulin-Like Growth Factor II / Q000008:administration & dosage*; D051379:Mice; D008810:Mice, Inbred C57BL; D010958:Plasminogen / Q000008:administration & dosage*; D011247:Pregnancy; D018873:Pregnancy Rate; D014568:Urokinase-Type Plasminogen Activator / Q000008:administration & dosage*
chemical_list: D003470:Culture Media; D007335:Insulin-Like Growth Factor II; D010958:Plasminogen; D014568:Urokinase-Type Plasminogen Activator