Preliminary results of the use of tumor cells of a dog to condition bovine embryo in vitro culture medium

MIGUEL MORENO MILLÁN; JESUS OCAÑA QUERO; LEONARDO DE LUCA; MANUEL HIDALGO; JESUS DORADO; SEBASTIÁN DEMYDA PEYRÁS

Córdoba

Congreso; 36th International Embryo Transfer Society Anual Meeting; 2010

IETS (International Embryo Transfer Society)

The aim of this study was to analyze the response of bovine embryos fertilized in vitro to a tumor dog cell- conditioned TCM-199 culture medium used in the development stage. Two hundred ninety-five oocytes obtained from slaughterhouse ovaries were cultured in groups of 20 cells at 39◦C in TCM-199 medium droplets with 20% estrous cow serum (ECS) for 24 h in a humid atmosphere containing 5% CO2. After incubation, these oocytes were fertilized with a 1×106 sperm cell dose, previously capacitated by a 1-h incubation in SP-TALP medium under usual conditions, by incubation in small petri dishes with 1 mL of Fert-TALP medium for 24 h under the conditions described previously. Alleged fertilized oocytes were randomly separated into 2 groups. A control group (n=144) was cultured in TCM-199 medium with 10% fetal calf serum (FCS) and the other (n=151) was cultured in TCM-199 medium previously conditioned by incubation for 72 h with 1×106 cells of a complex adenocarcinoma grade 1 obtained from a 6-year-old boxer dog and sterilized by filtration with 0.2-μm membrane. After 72 h of culture, the zygotes were harvested and prepared for a quick microscope observation. Alleged zygotes were classified as ?not fertilized,? ?divided,? or ?degenerated.? Statistical differences (P