Establishment of An Advanced Chemically Defined Medium for Early Embryos Derived from In Vitro Matured and Fertilized Bovine Oocytes
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概要
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A chemically defined medium would be useful for analyzing promoters or inhibitors in in vitro culture (IVC) of bovine embryos. However, an IVC system for bovine embryos in a chemically defined medium has not been fully established. The present study was carried out to establish an advanced chemically defined medium for bovine embryos that supports a high rate of embryo development to the blastocyst stage. In the first experiment, we examined the effects of addition of Medium RD (RPMI1640 and Dulbeccos MEM, 1:1 v/v) to mKSOM/aa on developmental competence. The addition of 10% RD to mKSOM/aa with BSA improved the rate of development to the blastocyst stage; however, 10% RD-mKSOM/aa with PVP, which is a chemically defined medium, caused a reduction in the percentage of hatching blastocysts. In the second experiment, embryos were cultured in the chemically defined medium of 10% RD-mKSOM/aa containing 11.7, 23.4, 46.8, 70.2 or 96.8 μM inositol. Inositol at the concentration of 70.2 μM improved the rate of development to the hatching blastocyst stage. In the third experiment, the optimal RD concentration in the IVC medium was evaluated. Embryos were cultured in the chemically defined medium supplemented with 10, 20 or 30% (v/v) RD. The rate of development to the blastocyst stage was highest with 20% RD. In the fourth experiment, the effects of N-acetylglucosamine (GlcNAc) as an IVC medium supplement on developmental competence were examined. The rate of development to the blastocyst stage with 1.0 mM GlcNAc was significantly higher than that without GlcNAc, but the rate of development with 1.2 mM GlcNAc was not different from that without GlcNAc. We also evaluated the ability of blastocysts produced in RD-mKSOM/aa to develop to normal calves after being transferred into recipients. Ten of the 16 recipients became pregnant, with 9 delivering normal calves. These results indicate that 20% RD-mKSOM/aa containing 70.2 μM myo-inositol and 1 mM GlcNAc is useful as a chemically defined medium for IVC of bovine embryos.
- 2011-12-01
著者
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Fukuda Yoshinori
School Of Veterinary Medicine And Animal Sciences Kitasato University
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Momozawa Kenji
School Of Veterinary Medicine And Animal Sciences Kitasato University
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Fukuda Yoshinori
School Of Veterinary Medicine Kitasato University
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Momozawa Kenji
School Of Veterinary Medicine Kitasato University
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MOMOZAWA Kenji
School of Veterinary Medicine and Animal Science, Kitasato University
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FUKUDA Yoshinori
School of Veterinary Medicine and Animal Science, Kitasato University
関連論文
- In Vitro Production of Blastocyst and Embryo Transfer in Mammals
- 受精能獲得精子を用いたウシ体外受精では受精培地へのカフェイン添加は不必要
- Establishment of An Advanced Chemically Defined Medium for Early Embryos Derived from In Vitro Matured and Fertilized Bovine Oocytes