To increase the success rate of the in vitro production (IVP) system of embryos in buffalo, it is necessary to improve parameters. This field requires more research since its results are low compared to cattle. In this study carried out in Colombia, a modification of the methodology to produce buffalo IVP embryos is described, and 24 non-lactating females, aged 4 to 11 years old were included. Follicular wave was synchronised by ablation of the follicles within the ovary, seven days later all follicles were aspirated to obtain oocytes for maturation, fertilisation and embryo production. For all culture media used, animal protein was avoided. Culture media were supplemented with growth factors and other molecules associated with embryonic development. Special care was taken in adding supplements to protect the zygotes against the harmful effect of oxygen free radicals in the culture medium. Two aspiration series were performed 15 days apart. The average number of oocytes obtained by aspiration was 11.33, the number of blastocysts was 3.05, and the obtained blastocyst production rate (36.2%) was higher than literature reports for this species; we hypothesise that this increase is associated with the supplements added. It was possible to observe a female that produced 50 oocytes. Subsequent research is needed to evaluate the effect of the medium on embryo quality and pregnancy rates. A cost analysis of the new proposed medium is required, since all supplements must be imported and handled with a cold chain of up to -80oC, making it more expensive and restricting widespread use. Finally, it is shown that with three embryos per aspiration, at least one pregnancy can be successful after the aspiration, which is one of the objectives of the IVP system.
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