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In Vitro Maturation of Porcine Oocytes in a Dry Incubator without CO2 Gas Supplement
The present study was conducted to develop a simple method for porcine oocyte maturation without CO2 regulation. In experiment 1, we evaluated that the effect of CO2 non-supplement on porcine oocyte maturation. Cumulusoocyte complexes (COCs) were collected from 2～6 mm follicles and divided into three groups (Control, tube-CO2, and tube-non-CO2). For control, COCs were cultured in 4-well multidish in a CO2 incubator. For tube-CO2, COCs were cultured in a round-bottom tube in a CO2 incubator, and for tube-non-CO2, COCs were cultured in a round-bottom tube sealed tightly without CO2 supplement in a dry incubator. The proportion of oocytes reached to metaphase II (M-II) was not significantly different among three groups (87.9% to 91.4%). In experiment 2, we evaluated the effect of CO2 non-supplement during oocyte maturation on development of embryos. Oocytes with a polar body were divided into two groups (Control and tube-non-CO2) and applied 1.1 kV/cm or 1.2 kV/cm voltages for parthenogenetic activation. After activation, embryos were cultured for 6 days and examined the development. The proportion of embryos cleaved was not significantly different among treatment (86.3% to 91.5%). The proportion of embryo reached to blastocyst stage was not significantly different among treatment (13.9% to 25.2%). The cell number of blastocysts was not significantly different among treatment (29.0 to 32.4). In conclusion, oocytes cultured in a dry incubator without CO2 supplement have enough competence to development after parthenogenetic activation. These results would be useful for transporting oocytes or embryos a long distance.
