Meiotic response of in vitro matured canine oocytes under different proteins and heterologous hormone supplementation.

摘要: Contents The impact of TCM-199 supplemented with different proteins and heterologous hormones on the in vitro maturation (IVM) rate bitch oocytes was evaluated by nuclear staining under fluorescence microscopy. Oocytes were recovered slicing ovaries from bitches presented at various stages oestrous cycle to ovariohysterectomy. The basic culture medium 25 mM Hepes/l, 10% heat-inactivated cow serum (ECS), 50 μg/ml gentamicin, 2.2 mg/ml sodium bicarbonate 22-μg/ml pyruvic acid, 1.0-μg/ml oestradiol (E 8875; Sigma), 0.5-μg/ml follicle-stimulating hormone (FSH) (Folltropin-V; Vetrepharm Inc., Ontario, Canada) 0.03 IU/ml human gonadotropin (hCG) (Profasi HP; Serono, Aubonne, Switzerland). Oocytes distributed randomly between (control) corresponding experimental treatment. Hormone treatments were: cultured in; (1) without FSH, (2) control 20 μg/ml oestradiol, or (3) 1 μg/ml somatotropin (hST; Humatrope, Lilly, Saint Cloud, France). The second experiment consisted 0.4% (w/v) bovine albumin (BSA, fraction V; Gibco Grand Island, NY, USA) instead ECS, inactivated (EBS) ECS. 100 μl droplets (up 25 per drop) mineral oil 37°C a 100% humidified atmosphere containing 5% CO2 air. After 72 h IVM, highest rates (p < 0.05) meiotic resumption achieved BSA supplementation. A positive influence metaphase II (MII) acquisition observed hST supplement. EBS supplementation did not develop MII stage. The results this study show that protein supplements tested promote final steps IVM oocytes.