title: Offspring from intracytoplasmic sperm injection of aged mouse oocytes treated with caffeine or MG132.
abstract: Postovulatory mammalian oocytes age significantly in culture. B6D2F1 or ICR strain mouse oocytes were collected 16 h after hCG injection and then cultured for up to 40 h post hCG at 37 °C under 5% CO(2) in air. After intracytoplasmic sperm injection (ICSI), B6D2F1 and ICR oocytes lost full-term developmental potential by 30 h and 26 h after hCG administration, respectively. However, using supplementation with 10 mM caffeine or 1-5 μM of MG132, we could obtain live offspring from oocytes at 34 h (BDF1, 5%-21%) or 28 h (ICR, 5%-18%), whereas none were obtained from untreated aged oocytes. Caffeine maintained normal meiotic spindle morphology, whereas MG132 maintained maturation-promoting factor activity. These treatments did not affect the potential of fresh oocytes for fertilization and subsequent development. Thus, it should be safe to use these chemicals in routine in vitro fertilization and offspring could be generated by ICSI of aged fertilization failed oocytes.
authors.affiliation: Laboratory for Genomic Reprogramming, RIKEN Center for Developmental Biology, Kobe 650-0047, Japan. onimarumaru@hotmail.com
authors.forename: Tetsuo
authors.initials: T
authors.lastname: Ono
authors.forename: Eiji
authors.initials: E
authors.lastname: Mizutani
authors.forename: Chong
authors.initials: C
authors.lastname: Li
authors.forename: Kazuo
authors.initials: K
authors.lastname: Yamagata
authors.forename: Teruhiko
authors.initials: T
authors.lastname: Wakayama
mesh_terms: D000367:Age Factors; D000818:Animals; D002110:Caffeine / Q000494:pharmacology*; D018929:Cell Culture Techniques; D016922:Cellular Senescence / Q000187:drug effects*; D006063:Chorionic Gonadotropin; D047108:Embryonic Development / Q000187:drug effects*; D005307:Fertilization in Vitro / Q000379:methods*; D007976:Leupeptins / Q000494:pharmacology*; D016200:Maturation-Promoting Factor / Q000378:metabolism; D051379:Mice; D008813:Mice, Inbred ICR; D009865:Oocytes / Q000187:drug effects*; D020554:Sperm Injections, Intracytoplasmic; D008941:Spindle Apparatus / Q000187:drug effects
chemical_list: D006063:Chorionic Gonadotropin; D007976:Leupeptins; D002110:Caffeine; D016200:Maturation-Promoting Factor; C072553:benzyloxycarbonylleucyl-leucyl-leucine aldehyde