The effect of pre-maturation culture using phosphodiesterase type 3 inhibitor and insulin, transferrin and selenium on nuclear and cytoplasmic maturation of bovine oocytes.
This study aims to evaluate if a pre-maturation culture (PMC) using cilostamide as a meiotic inhibitor in combination with insulin, transferrin and selenium (ITS) for 8 or 24 h increases in vitro embryo production. To evaluate the effects of PMC on embryo development, cleavage rate, blastocyst rate, embryo size and total cell number were determined. When cilostamide (20 μM) was used in PMC for 8 or 24 h, 98% of oocytes were maintained in germinal vesicles. Although the majority of oocytes resumed meiosis after meiotic arrest, the cleavage and blastocyst rates were lower than the control (P 0.05) to the control. The deleterious effect of 20 μM cilostamide treatment for 24 h on a PMC was confirmed by lower cumulus cell viability, determined by trypan blue staining, in that group compared with the other groups. A lower concentration (10 μM) and shorter exposure time (8 h) minimized that effect but did not improve embryo production. More studies should be performed to determine the best concentration and the arresting period to increase oocyte competence and embryo development.
School of Agriculture and Veterinary Medicine,University of Brasilia,Brasília-DF,Brazil.
A L S
AL
Guimarães
Institute of Biology,University of Brasília,Brasília-DF,Brazil.
S A
SA
Pereira
School of Veterinary Medicine,University of Uberlândia,Uberlândia-MG,Brazil.
N R
NR
Kussano
Parque Estação Biológica,Final Av. W5/N,Prédio PBI,70770-900,Brasília-DF,Brazil.
M A N
MA
Dode
D000818:Animals; D001755:Blastocyst / Q000187:drug effects / Q000502:physiology; D002417:Cattle; D002467:Cell Nucleus / Q000187:drug effects / Q000378:metabolism; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D054885:Cumulus Cells / Q000187:drug effects; D003593:Cytoplasm / Q000187:drug effects / Q000378:metabolism; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D059471:In Vitro Oocyte Maturation Techniques / Q000379:methods / Q000662:veterinary; D007328:Insulin / Q000494:pharmacology*; D008297:Male; D008540:Meiosis / Q000187:drug effects; D009865:Oocytes / Q000166:cytology / Q000187:drug effects* / Q000502:physiology; D058987:Phosphodiesterase 3 Inhibitors / Q000494:pharmacology*; D015363:Quinolones / Q000494:pharmacology; D012643:Selenium / Q000494:pharmacology*; D013997:Time Factors; D014168:Transferrin / Q000494:pharmacology*
D007328:Insulin; D058987:Phosphodiesterase 3 Inhibitors; D015363:Quinolones; D014168:Transferrin; C021294:cilostamide; D012643:Selenium
Bovine oocyte; Cilostamide; Embryo; Meiotic arrest; Pre-maturation culture