title: Effects of vitrification solutions and equilibration times on the morphology of cynomolgus ovarian tissues.
abstract: This study assessed the effects of vitrification solutions and equilibration times on morphology of cynomolgus ovarian tissues. Ovarian cortical sections (0.1-0.2 cm thickness) of seven cynomolgus monkeys were randomly allocated to either a control group or one of six vitrification groups. Ovarian tissue sections were vitrified ultra-rapidly by placing them directly into liquid nitrogen using two different vitrification solutions (VSEGP: 5.64 mol/l ethylene glycol+5% (w/v) polyvinylpyrrolidone+0.5 mol/l sucrose; and VSED: 3.22 mol/l ethylene glycol+2.56 mol/l dimethylsulphoxide+0.5 mol/l sucrose) after three different exposure times (5-20 min). After warming, follicle morphology was analysed using light and transmission electron microscopy. The proportion of morphologically normal follicles vitrified using VSED after a 5-min exposure was lower (P<0.05) than those vitrified by other conditions. The proportion of normally structured mitochondria in oocytes of preantral follicles vitrified after a 5-min exposure to VSED (56%) was lower (P<0.01) than those vitrified by other conditions (78-88%). Following tissue vitrification with VSED, the surface ratio of lysosome was increased compared with non-vitrified oocytes (1.64% versus 1.11%; P<0.05). These results indicate that VSEGP can support the morphology of vitrified preantral follicles and oocytes.
authors.affiliation: IVF Namba Clinic, Osaka, Osaka 550-0015, Japan. hashimoto@ivfnamba.com
authors.forename: Shu
authors.initials: S
authors.lastname: Hashimoto
authors.forename: Nao
authors.initials: N
authors.lastname: Suzuki
authors.forename: Masaya
authors.initials: M
authors.lastname: Yamanaka
authors.forename: Yoshihiko
authors.initials: Y
authors.lastname: Hosoi
authors.forename: Bunpei
authors.initials: B
authors.lastname: Ishizuka
authors.forename: Yoshiharu
authors.initials: Y
authors.lastname: Morimoto
mesh_terms: D000818:Animals; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D019855:Ethylene Glycol / Q000494:pharmacology*; D005260:Female; D008247:Lysosomes / Q000187:drug effects; D008252:Macaca fascicularis; D046529:Microscopy, Electron, Transmission; D008928:Mitochondria / Q000187:drug effects; D009865:Oocytes / Q000187:drug effects / Q000648:ultrastructure*; D006080:Ovarian Follicle / Q000187:drug effects / Q000648:ultrastructure*; D010053:Ovary / Q000187:drug effects / Q000648:ultrastructure*; D011205:Povidone / Q000494:pharmacology*; D012996:Solutions / Q000494:pharmacology; D013395:Sucrose / Q000494:pharmacology*; D058989:Vitrification
chemical_list: D012996:Solutions; D013395:Sucrose; D019855:Ethylene Glycol; D011205:Povidone; D004121:Dimethyl Sulfoxide