Effect of culture media on embryo development from prepubertal goat IVM-IVF oocytes.
Experiments were carried out to develop and improve in vitro culture systems for IVM-IVF prepubertal goat oocytes. Cumulus oocyte complexes (COC) were obtained by slicing ovaries from slaughtered prepubertal goats. Oocytes were matured in TCM-199 supplemented with 20% estrous goat serum (EGS) + 10 micrograms/mL FSH + 10 micrograms/mL LH + 1 microgram/mL estradiol 17 beta for 27 h at 38.5 degrees C in 5% CO2 in air. Matured oocytes were placed in drops of TALP- fert medium supplemented with hypotaurine (1 microgram/mL) and inseminated with freshly ejaculated spermatozoa following capacitation as described by Younis et al. (69) but with 100 micrograms/mL heparin. At 24 h post insemination the ova were transferred to various in vitro culture media, and early embryo development was evaluated until Day 8 post insemination. Specifically, in the studies described here, we have compared the effects of (Experiment 1) co-culture systems using oviductal ephitelial cells (OEC) and cumulus cells (CC), both caprine and bovine; (Experiment 2) the presence of serum and/or OEC; (Experiment 3) 4 culture media (TCM199, Ham's F10, CZB abd SOF) for co-culture with OEC; and (Experiment 4) conditioned medium with OEC. In Experiment 1, the percentage of morulae plus blastocysts was higher in culture with OEC, both caprine and bovine (15.1 and 14.8%, respectively) than with CC (4.1 and 6.7%, respectively). In Experiment 2, the OEC with EGS did not improve the percentage of morulae and blastocysts obtained with OEC alone (14.3 and 23.1% respectively). In Experiment 3, this percentage was higher using OEC with TCM-199 compared to CZB medium (21.3 and 12.3%, respectively) and in Experiment 4, the results were 3.7, 11.2 and 21.3% for TCM-199 without cells, Conditioned Medium and co-culture with OEC, respectively.
Izquierdo
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Departament de Patologia i de Producció Animal Universitat Autònoma de Barcelona, Bellaterra, Spain.
Villamediana
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D000818:Animals; D001755:Blastocyst / Q000166:cytology / Q000502:physiology*; D002417:Cattle; D002478:Cells, Cultured; D018920:Coculture Techniques; D003470:Culture Media; D005314:Embryonic and Fetal Development / Q000502:physiology*; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D005307:Fertilization in Vitro / Q000379:methods / Q000662:veterinary; D006041:Goats; D006493:Heparin / Q000494:pharmacology; D008297:Male; D009028:Morula / Q000166:cytology / Q000502:physiology*; D018587:Oocyte Donation / Q000662:veterinary; D009865:Oocytes / Q000166:cytology* / Q000502:physiology; D010053:Ovary / Q000166:cytology; D012741:Sexual Maturation; D013094:Spermatozoa / Q000187:drug effects / Q000502:physiology; D013654:Taurine / Q000031:analogs & derivatives
D003470:Culture Media; D013654:Taurine; C003949:hypotaurine; D006493:Heparin