title: In vitro maturation of pig oocytes with different media, hormone and meiosis inhibitors.
abstract: This study evaluated in vitro maturation of pig oocytes in two maturation media (TCM199 and NCSU23) supplemented with 10% porcine follicular fluid (pFF) or 0.1% polyvinyl alcohol (PVA) and four hormonal treatments. The best media was then used to evaluate the effect of reversible meiosis inhibitors cycloheximide (5 microgram/ml) [DOSAGE ERROR CORRECTED]and butyrolactone I (12.5M) on the maturation of pig oocytes was evaluated. After maturation for 44 h, the oocytes were fixed, stained, and examined under epifluorescence microscopy. The comparison of the proportion of oocytes in metaphase II revealed that hormonal treatment 2(incubation for 22 h - 10 ng EGF/ml, 10 IU hCG/ml and 10 IU eCG/ml, followed by incubation for 22 h - 10 ng EGF/ml) presented higher repeatability percentages: TCM+ PVA (54.5% - 61/112); TCM+ pFF (65.0% - 63/97);NCSU23 + PVA (54.6% - 65/119), and NCSU23 + pFF (58.1% - 61/105). The comparison of maturation media showed that TCM199 presented more constant results than NCSU23. Regarding supplementation with pFF or PVA, TCM199 with pFF presented better results. The comparison between butyrolactone I and cycloheximide demonstrated that both drugs effectively inhibited meiosis; however, only cycloheximide presented metaphase II percentages similar to the control (70.29% and 75.49%, respectively). In conclusion, it is recommended the use of TCM199 medium supplemented with pFF and hormonal treatment with 10 ng EGF/ml, 10 UI hCG/mland 10 UI eCG/ml during the first 22 h and more 22 h with 10 ng EGF/ml for the pig oocytes maturation. Butyrolactone I and cycloheximide effectively arrested/resumpted maturation; however, the oocytes percentages in metaphase II was the same for both cycloheximide and the control groups.
authors.affiliation: Department of Animal Reproduction, Faculty of Veterinary Medicine, University of São Paulo, Brazil. mgroke@usp.br
authors.forename: Mariana Groke
authors.initials: MG
authors.lastname: Marques
authors.forename: Alessandra Corallo
authors.initials: AC
authors.lastname: Nicacio
authors.forename: Viviane Purri
authors.initials: VP
authors.lastname: de Oliveira
authors.forename: Anibal Ballarotti
authors.initials: AB
authors.lastname: Nascimento
authors.forename: Heloisa Vasconcelos Amaral
authors.initials: HV
authors.lastname: Caetano
authors.forename: Camilla Mota
authors.initials: CM
authors.lastname: Mendes
authors.forename: Marco Roberto Bourg
authors.initials: MR
authors.lastname: Mello
authors.forename: Marcella Pecora
authors.initials: MP
authors.lastname: Milazzotto
authors.forename: Mayra Elena Ortiz D'Avila
authors.initials: ME
authors.lastname: Assumpção
authors.forename: José Antônio
authors.initials: JA
authors.lastname: Visintin
mesh_terms: D015107:4-Butyrolactone / Q000031:analogs & derivatives / Q000494:pharmacology; D000818:Animals; D006063:Chorionic Gonadotropin / Q000494:pharmacology; D003470:Culture Media / Q000737:chemistry*; D003513:Cycloheximide / Q000494:pharmacology; D004305:Dose-Response Relationship, Drug; D046149:Embryo Culture Techniques / Q000379:methods / Q000662:veterinary*; D004815:Epidermal Growth Factor / Q000494:pharmacology; D005260:Female; D005307:Fertilization in Vitro / Q000662:veterinary; D015571:Follicular Fluid / Q000502:physiology; D006064:Gonadotropins, Equine / Q000494:pharmacology; D008540:Meiosis / Q000187:drug effects*; D008856:Microscopy, Fluorescence / Q000662:veterinary; D009865:Oocytes / Q000187:drug effects / Q000254:growth & development; D011142:Polyvinyl Alcohol / Q000494:pharmacology; D011500:Protein Synthesis Inhibitors / Q000494:pharmacology; D013552:Swine; D013997:Time Factors
chemical_list: D006063:Chorionic Gonadotropin; D003470:Culture Media; D006064:Gonadotropins, Equine; D011500:Protein Synthesis Inhibitors; D004815:Epidermal Growth Factor; C082868:butyrolactone I; D011142:Polyvinyl Alcohol; D003513:Cycloheximide; D015107:4-Butyrolactone