Effects of oxygen, CO2/pH and medium on the in vitro development of individually cultured porcine one- and two-cell embryos

The gas atmosphere and medium composition are critical factors in the in vitro development of one- and two-cell embryos of several species. The present study evaluated the effect of different O-2/CO2 concentrations (2/5, 2/10, 5/2.5, 5/5, 5/10, 10/10 and 21/5) on pig one- and two-cell embryo development. The embryos were individually cultured, for 6 days at 39 degrees C in a medium rich in bicarbonate and glutamine and containing pyruvate and lactate but lacking glucose. When the CO2, levels increased from 2.5% to 10%, the pH of the medium decreased from 8.2. to 7.5 and the development of the embryos was affected, but this depended mainly on the O-2 levels. Pig embryo development was inhibited by 2 and 21% O-2 levels. The optimum level for pig embryo development was 5% O-2 and 5% CO2, whatever the criteria used to evaluate embryo development. At these optimal levels, the mean number of cells per embryo was 26 +/- 1.7 (Is mean +/- SE), and 50% of the one- and two-cell embryos developed to blastocysts. The substitution of 0.5% bovine serum albumin (BSA) in the medium by 0.3% polyvinyl-pyrrolidone (PVP) significantly decreased the one- and two-cell embryo development. When the calcium and chloride contents of the medium with PVP were reduced, however, the embryo development was similar to that observed in the medium containing BSA. Pig embryo development in vitro was found to be optimal under an atmosphere of 5% O-2, and 5% CO2 and PVP could replace BSA as the high molecular weight supplement.