Effect of Different Culture Conditions on In vitro Development and Quality of Buffalo Embryos

Abstract

This work was carried out to evaluate the quality and the developmental competence of the in vitro buffalo embryos produced under different culture conditions. Oocytes were matured in TCM 199 for 24 h at 38.5°C and 5% CO 2 . After maturation, oocytes were in vitro fertilized by frozen thawed sperms capacitated in vitro by caffeine at final concentration 2 million sperm /ml. The sperm-oocytes were incubated in TALP medium for 24 hrs. Fertilized oocytes were cultured in Cell free media (CR1aa) or with cumulus or oviductal cell monolayers (Experiment 1), CR1aa with addition of (10, 20, 50 and 100 µg/mL) lactoferrin (Experiment 2) inside CO 2 incubator with 5% CO 2 at 38°C. The cleavage, morula, blastocyst rates were recorded and embryo cell number and mitochondrial function were evaluated. Addition of bovine oviductal cell monolayer (BOCM) to culture media significantly increased the morula and the blastocyst compared with cell free media and media with cumulus cell monolayer and also significantly increased the cell number and the mitochondrial function of in vitro produced buffalo embryos. Addition of lactoferrin by concentration 50µg/ml to culture media resulted in a significant increase in rates of cleavage; morula and blastocyst compared with control or other concentrations and also resulted in an increase in the cell number and the mitochondrial function of in vitro produced buffalo embryos. It could be concluded that, addition of oviductal cells (monolayer) or lactoferrin to culture medium significantly improved the quality of in vitro produced buffalo embryos.