Regulation of culture media composition and conditions on embryonic sex ratio and development in in vitro produced buffalo embryos

The production of female offspring is vital for maximizing milk yield in the dairy industry, making embryo sexing an essential tool in assisted reproductive technologies (ART). This study aimed to investigate the effect of culture media composition on the sex ratio of in vitro produced buffalo embryos. Ovaries were collected from slaughterhouse buffaloes, and oocytes were retrieved and matured in vitro for 24 hours. Following in vitro fertilization (IVF), presumptive zygotes were cultured in either a sequential Synthetic Oviduct Fluid (SOF) medium—lacking glucose in early stages and supplemented with glucose in later stages—or in a single-step Potassium Simplex Optimized Medium (KSOM). After six days of culture, morula-stage embryos were harvested, and sex determination was performed using polymerase chain reaction (PCR). Analysis of the resulting sex ratios revealed that culture media composition influenced the proportion of male and female embryos. These findings underscore the importance of optimizing culture conditions to favor the production of female embryos in buffalo, with potential applications in dairy herd management.