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KMID : 1023620120360030141
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Reproductive and Developmental Biology
2012 Volume.36 No. 3 p.141 ~ p.145
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In Vitro Maturation of Porcine Oocytes in a Dry Incubator without CO₂ Gas Supplement
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Park Kwang-Wook
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Abstract
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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.
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KEYWORD
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Pig, Oocytes, In Vitro Maturation, Dry Incubator
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