Pporcine reproductive biology has gained more interest recently due to an increase in pig meat requests; demand to enhance and preserve the good genetic material of the animal, along with improvement of animal genetic transportation biosecurity made the reproductive biotechnology create more suitable ways to achieve this destination, in vitro embryo production (IVEP) is one of this ways. Norsvin SA is a research and development-based company looking for optimization of the IVEP process. Porcine IVEP is more challenging than most domestic animals, due to gamete sensitivity. On the other hand, for the research purposes, due to the utilization of oocytes from the slaughterhouse, assessment, and control of the oocyte quality is even more out of reach, therefore, in this study, the main focus was on sperm cell quality (motility criteria, viability, acrosome reactivity) assessment by CASA and flocytometry. This study tried to find standardization for sperm selection by examining two different percoll batches and storage time. Furthermore, it was proven by prior studies that sperm concentration in IVF plays a crucial role in the fertilization and blastocyst rate; therefore after sperm selection by percoll discontinuous gradient, and oocyte selection and maturation two different sperm concentrations per COC were applied: 1’000:1 ratio (normal) and 2’000:1 ratio (double). The data analysis after culturing potential zygote revealed no significant difference between the two treatments at cleavage and blastocyst rate (day 6 and 7) (n= 935; P>0.05). However, in polyspermy assessment of a total of 120 cells, 0.0%, and 13.4% polyspermy were observed in normal and double sperm concentrations respectively. In a conclusion, it was proven that the 1’000:1 ratio of sperm cell/COC seemed to be a more suitable ratio for IVF.
Keywords: in vitro embryo production (IVEP), in vitro fertilization (IVF), sperm selection, cleavage and blastocyst rate, polyspermy