title: Efficient production of live offspring from mouse oocytes vitrified with a novel cryoprotective agent, carboxylated ε-poly-L-lysine.
abstract: In cryopreservation of mammalian germ cells, unfertilized oocytes are one of the most available stages because these cryopreserved oocytes can be used for assisted reproductive technologies, including in vitro fertilization (IVF) and intracytoplasmic sperm injection. However, it has been generally reported that the fertility and developmental ability of the oocytes are reduced by cryopreservation. Therefore further improvement will be required. Very recently, a new cryoprotective agent (CPA), called as carboxylated ε-poly-L-lysine (COOH-PLL), has been developed to reduce physical and physiological damage by cryopreservation in mammalian stem cells. However, it is unclear the effect of COOH-PLL on fertility and developmental ability of vitrified oocytes. In this study, we used COOH-PLL as a CPA with ethylene glycol (EG) for vitrification of mouse oocytes. Cumulus-oocyte complexes (COCs) were collected from ICR mice and then vitrified with Cryotop using different concentration of COOH-PLL and EG. A combined treatment with COOH-PLL and EG showed high survival rate (more than 90%) of vitrified-warmed COCs after in vitro fertilization. In addition, the fertility and developmental ability of COCs vitrified with E20P10 [EG 20% (v/v) and COOH-PLL 10% (w/v)] or E15P15 group (EG 15% and COOH-PLL 15%) were significantly higher than those with E10P20 (EG10% and COOH-PLL 20%) or P30 group (PLL30%). The vitrified COCs in E20P10 group developed to term at a high success rate (46.2%) and it was significantly higher than that in control (E30) group (34.8%). Our present study demonstrated for the first time that COOH-PLL is effective for vitrification of mouse oocytes.
authors.affiliation: Laboratory of Animal Reproduction, School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan.
authors.forename: Hitomi
authors.initials: H
authors.lastname: Watanabe
authors.affiliation: Laboratory of Animal Reproduction, School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan.
authors.forename: Natsuki
authors.initials: N
authors.lastname: Kohaya
authors.affiliation: Laboratory of Animal Reproduction, School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan.
authors.forename: Maki
authors.initials: M
authors.lastname: Kamoshita
authors.affiliation: Graduate School of Veterinary Sciences, Azabu University, Sagamihara, Kanagawa, Japan.
authors.forename: Katsuyoshi
authors.initials: K
authors.lastname: Fujiwara
authors.affiliation: Japan Advanced Institute of Science and Technology, Nomi, Ishikawa, Japan.
authors.forename: Kazuaki
authors.initials: K
authors.lastname: Matsumura
authors.affiliation: Center for Fiber and Textile Science, Kyoto Institute of Technology, Matsugasaki, Kyoto, Japan.
authors.forename: Suong-Hyu
authors.initials: SH
authors.lastname: Hyon
authors.affiliation: Laboratory of Animal Reproduction, School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan ; Graduate School of Veterinary Sciences, Azabu University, Sagamihara, Kanagawa, Japan.
authors.forename: Junya
authors.initials: J
authors.lastname: Ito
authors.affiliation: Laboratory of Animal Reproduction, School of Veterinary Medicine, Azabu University, Sagamihara, Kanagawa, Japan ; Graduate School of Veterinary Sciences, Azabu University, Sagamihara, Kanagawa, Japan.
authors.forename: Naomi
authors.initials: N
authors.lastname: Kashiwazaki
mesh_terms: D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000737:chemistry* / Q000494:pharmacology*; D019855:Ethylene Glycol / Q000737:chemistry; D005260:Female; D005298:Fertility / Q000187:drug effects; D005307:Fertilization in Vitro; D051379:Mice; D008813:Mice, Inbred ICR; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D011107:Polylysine / Q000737:chemistry* / Q000494:pharmacology*; D058989:Vitrification / Q000187:drug effects
chemical_list: D003451:Cryoprotective Agents; D011107:Polylysine; D019855:Ethylene Glycol