title: Live offspring from cryopreserved embryos following in vitro growth, maturation and fertilization of oocytes derived from preantral follicles in mice.
abstract: This study was undertaken to examine pre- and postimplantation developmental potency of cryopreserved embryos that had undergone in vitro growth (IVG), maturation (IVM) and fertilization (IVF) of oocytes from the preantral follicle stage. An oocyte culture system for IVG and IVM was used in oocyte-granulosa cell complexes (OGCs) derived from preantral follicles in 12-day-old mice. The rate of oocyte maturation was improved by the addition of gonadotropins (FSH/LH) and cytokines (IGF-I/SCF) to culture medium for IVG. During culture for IVG, estradiol-17β and progesterone concentrations increased progressively to the latter period of culture. This culture system enabled IVG, IVM, IVF and pre- and postimplantation development. From 90 cryopreserved 2-cell stage embryos transferred into recipients after warming, 10 live pups were produced. Cryopreservation of embryos by vitrification at the 2-cell stage showed no harmful effect on development to the blastocyst stage or on the cell numbers of the inner cell mass (ICM) and trophectoderm (TE). This study demonstrated that embryos derived from oocytes grown in vitro have tolerance for vitrification and competence to develop to term after warming. This IVG-IVM-IVF technology combined with embryo cryopreservation might be useful for assisted reproduction in mice.
authors.affiliation: Department of Bioproduction Technology, Junior College of Tokyo University of Agriculture, Tokyo 156-8502, Japan. motohashi@ncnp.go.jp
authors.forename: Hideyuki H
authors.initials: HH
authors.lastname: Motohashi
authors.forename: Tadashi
authors.initials: T
authors.lastname: Sankai
authors.forename: Hidemi
authors.initials: H
authors.lastname: Kada
mesh_terms: D000818:Animals; D001755:Blastocyst / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003470:Culture Media; D004958:Estradiol / Q000096:biosynthesis; D005260:Female; D005307:Fertilization in Vitro / Q000187:drug effects / Q000379:methods; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D006107:Granulosa Cells / Q000187:drug effects; D007334:Insulin-Like Growth Factor I / Q000494:pharmacology; D050498:Live Birth; D007986:Luteinizing Hormone / Q000494:pharmacology; D051379:Mice; D008810:Mice, Inbred C57BL; D008808:Mice, Inbred CBA; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development*; D006080:Ovarian Follicle / Q000187:drug effects / Q000254:growth & development*; D011374:Progesterone / Q000096:biosynthesis; D019089:Stem Cell Factor / Q000494:pharmacology; D058989:Vitrification
chemical_list: D003470:Culture Media; D019089:Stem Cell Factor; C519530:insulin-like growth factor-1, mouse; D011374:Progesterone; D004958:Estradiol; D007334:Insulin-Like Growth Factor I; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone