Effects of supplementation of the maturation media with insulin on in vitro maturation and in vitro fertilization of bovine oocytes.
This study was carried out to determine the effects of supplementation of the maturation media with insulin on in vitro maturation and fertilization of bovine oocytes. In Experiment 1, cumulus-intact bovine oocytes were cultured in a maturation medium (TCM-199 containing 10% fetal calf serum, 0.02 U/ml follicular stimulating hormone and 1 microgram/ml estradiol-17 beta) with or without insulin supplementation (10 micrograms/ml). The maturation and fertilization rates of oocytes and subsequent embryonic development to the blastocyst stage were not affected by the treatment with insulin in the presence of serum and the hormones during the maturation period. In Experiment 2, to avoid the effects of serum and the hormones, a serum- and hormone-free maturation medium (TCM-199 containing 1 mg/ml polyvinyl alcohol) was used. In the absence of serum and hormones during the maturation period, the maturation rate was not affected by treatment with insulin, but the fertilization rate was improved. In Experiment 3, when denuded oocytes were inseminated together with cumulus cells cultured in serum- and hormone-free maturation medium supplemented with insulin, the fertilization rate was increased. These results demonstrate that the addition of insulin to the serum- and hormone-free maturation medium improves the fertilization rate of bovine oocytes in vitro, and suggest that insulin may stimulate the secretion of sperm capacitating agent (s) from cumulus cells.
Matsui
M
M
Department of Veterinary Clinical Science, Hokkaido University, Sapporo, Japan.
Takahashi
Y
Y
Hishinuma
M
M
Kanagawa
H
H
D000818:Animals; D002417:Cattle; D002454:Cell Differentiation / Q000187:drug effects / Q000502:physiology; D002478:Cells, Cultured; D003470:Culture Media / Q000032:analysis / Q000494:pharmacology*; D004958:Estradiol / Q000494:pharmacology; D005260:Female; D005298:Fertility / Q000187:drug effects / Q000502:physiology; D005306:Fertilization / Q000187:drug effects / Q000502:physiology; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary*; D005640:Follicle Stimulating Hormone / Q000494:pharmacology; D066298:In Vitro Techniques; D007328:Insulin / Q000032:analysis / Q000494:pharmacology*; D008297:Male; D009865:Oocytes / Q000166:cytology* / Q000187:drug effects* / Q000502:physiology; D011142:Polyvinyl Alcohol / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology; D013075:Sperm Capacitation / Q000187:drug effects / Q000502:physiology
D003470:Culture Media; D007328:Insulin; D012710:Serum Albumin, Bovine; D004958:Estradiol; D005640:Follicle Stimulating Hormone; D011142:Polyvinyl Alcohol