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A producao in vitro (PIV) de embrioes bovinos alcancada com vacas de matadouros ou de aspiracao folicular in vivo (OPU) e uma pratica cada vez mais difundida e a sua simplificacao poderia baixar os custos de producao. O objetivo desta pesquisa foi comparar a producao in vitro de embrioes em estufa com temperatura, umidade relativa e atmosfera controlada (controle), com tubos de poliestireno gaseificados e mantidos em banho-maria (tratamento). Oocitos obtidos de ovarios de vacas abatidas foram maturados in vitro em TCM- 199 modificado com 25mM de N-2-hidroxietilpiperazina-N -2-acido etanosulfonico (HEPES); 0,025mg/ml de piruvato de sodio, 0,01UI de rFSHh/ml, 0,5µg/ml de LHs e 10% de soro de vaca em estro. Na fecundacao in vitro utilizou-se Talp-Fert com 0,06mg/ml de albumina serica bovina, 0,022mg/ml de piruvato de sodio e 10µg/ml de heparina. O cultivo foi conduzido em placas de 4 pocos em SOF com 5% de soro de vaca em estro, 20µl/ml de aminoacidos essenciais e 10µl/ml de aminoacidos nao essenciais, sob oleo mineral, em estufa com atmosfera de 5% de CO2, umidade saturada e 39°C, por 9 dias. Na maturacao nao houve diferenca (P>0,05) entre o tratamento e o controle. Porem, a maturacao e a fecundacao ou somente a fecundacao in vitro em tubos mantidos em banho-maria nao demonstrou ser uma alternativa recomendada para a producao de embrioes bovinos. In vitro maturation and fertilization of bovine oocytes in tubes previously gasified kept in water bath Abstract In vitro bovine embryo production either obtained from oocytes of slaughtered cows or in vivo follicular aspiration (OPU) is a well-known technique and its simplification might reduce the cost of embryo production. The aim of this study was to compare the cleavage rate and embryo development of the in vitro production of bovine embryos using standard culture system (temperature, gas phase and controlled humidity) versus gasified polystyrene tubes kept in water bath. Oocytes obtained from ovaries of slaughtered cows were in vitro maturated in TCM- 199modified with 25 mM of N-2-hidroxyethylpiperazine-N-2-ethanosulfonic acid (HEPES); containing 0.01UI rFSHh/ml and 0.5µg/ml LHs, 0.025mg/ml sodium pyruvate and 10% estrous cow serum. The in vitro fertilization was carried out in Talp-Fert containing 0.06mg/ml BSA, 0.022mg/ml sodium pyruvate and 10µg/ml heparin. The culture was performed in SOF medium with 20µl/ml essential aminoacids, 10µl/ml, non-essential aminoacids and 5% estrous cow serum, with oil overlay, in 4 well dishes and incubated with 5% CO2, maximum humidity at 39°C, for 9 days. The results of this study showed no difference (P>0.05) between the treatment and control groups during the maturation process. However, the maturation and fertilization or only the fertilization in tubes do not represent a viable alternative for the in vitro production of bovine embryos.

MATURAÇÃO E FECUNDAÇÃO IN VITRO DE OÓCITOS BOVINOS EM TUBOS PREVIAMENTE GASEIFICADOS E MANTIDOS EM BANHO-MARIA