Effects of vitrification solutions and equilibration times on the morphology of cynomolgus ovarian tissues.
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.
Hashimoto
Shu
S
IVF Namba Clinic, Osaka, Osaka 550-0015, Japan. hashimoto@ivfnamba.com
Suzuki
Nao
N
Yamanaka
Masaya
M
Hosoi
Yoshihiko
Y
Ishizuka
Bunpei
B
Morimoto
Yoshiharu
Y
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
D012996:Solutions; D013395:Sucrose; D019855:Ethylene Glycol; D011205:Povidone; D004121:Dimethyl Sulfoxide