The effects of 17beta-estradiol and protein supplement on the response to purified and recombinant follicle stimulating hormone in bovine oocytes.
During in vitro maturation of bovine oocytes, effects of gonadotropins (FSH, LH) and growth factors such as epidermal growth factor (EGF) vary among studies. Now that we can use defined or semi-defined medium, it becomes possible to evaluate recombinant products to assess their roles. Therefore, this study was designed to evaluate the effect of purified porcine (pFSH) or recombinant human (r-hFSH; 5, 50, or 500 ng/ml) follicle stimulating hormone, luteinising hormone (LH; 50,500 or 5000 ng/ml) and epidermal growth factor (EGF; 5, 10, 30 or 50 ng/ml) on subsequent embryonic development of in vitro matured bovine oocytes. In addition, the presence of bovine serum albumin (BSA; 8 mg/ml) as a protein supplement during in vitro maturation (IVM) was studied. For all treatments, cumulus-oocyte complexes were matured in defined maturation medium consisting of synthetic oviduct fluid. Addition of LH to the maturation medium at all concentrations studied did not increase the proportion of oocytes developing to the morula and blastocyst stages. However, morula and blastocyst yield were improved (p < 0.05) after addition of EGF (30 ng/ml) as compared with maturation medium alone (29.3% vs 18.0%, respectively). Addition of r-hFSH to the maturation medium in the presence of 17beta-estradiol (E2) significantly (p < 0.0001) increased the morula and blastocyst rate compared with maturation medium alone (40.3% vs 19.3%, respectively). The presence of BSA alone during IVM significantly reduced the developmental competence of oocytes as reflected by the morula and blastocyst yield. These results demonstrate the essential role of FSH, EGF and E2 on the kinetics of nuclear and cytoplasmic maturation that are essential for the formation of an egg capable of fertilisation and development. Also, supplementation of r-hFSH and E2 during IVM under our conditions increases morula and blastocyst yield following in vitro fertilisation and in vitro culture in defined medium. Finally, the presence of BSA as the only protein supplement during IVM may be detrimental to oocyte maturation.
Ali
Atef
A
Centre de Recherche en Biologie de la Reproduction (CRBR), Department of Animal Science, Laval University, Quebec, Canada.
Sirard
Marc-André
MA
D000818:Animals; D001755:Blastocyst / Q000378:metabolism; D002417:Cattle; D002478:Cells, Cultured; D003470:Culture Media / Q000494:pharmacology*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D004958:Estradiol / Q000494:pharmacology*; D005260:Female; D005307:Fertilization in Vitro; D005640:Follicle Stimulating Hormone / Q000235:genetics / Q000494:pharmacology*; D007986:Luteinizing Hormone / Q000494:pharmacology; D008297:Male; D009028:Morula / Q000378:metabolism; D009865:Oocytes / Q000166:cytology*; D009866:Oogenesis / Q000502:physiology; D011994:Recombinant Proteins / Q000494:pharmacology; D012710:Serum Albumin, Bovine / Q000494:pharmacology*
D003470:Culture Media; D011994:Recombinant Proteins; D012710:Serum Albumin, Bovine; D004958:Estradiol; D004815:Epidermal Growth Factor; D007986:Luteinizing Hormone; D005640:Follicle Stimulating Hormone