Production of a Germ-line Chimera by Coculture of Zona-free Embryos with Frozen-thawed Embryonic Stem Cells
スポンサーリンク
概要
- 論文の詳細を見る
To improve chimera production efficiency, we examined whether frozen-thawed embry-onic stem (ES) cells would be useful in a coculture method. A3-1 ES cells were electroporated with the targeting vector pE 10.29NEO-TK. Exon 2 of the endothelin-1 gene in the ES cells was disrupted by homologous recombination. The homologous recombinant was cocultured at a density of 5.5 × 10<SUP>5</SUP> cells/ml with zona-free 8-cell to morula stage embryos in a coculture medium for 3-3.5 h. Cocultured embryos were transferred into Whittens medium and cultured overnight. Morula and blastocyst stage embryos were transferred into the recipients on day 2.5 of pseudopregnancy. The development rate to blastocyst after coculture and subsequent overnight cultivation was 62%. As a result of embryo transfer, four male chimeras were obtained. Two of these were identified as germ-line chimeras by a progeny test. These results indicate that frozen-thawed ES cells can be used in coculture to produce chimeras and contribute a germ-line.
- 日本繁殖生物学会の論文
著者
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KURIHARA Yukiko
Third Department of Internal Medicine, Faculty of Medicine, University of Tokyo
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AZUMA Sadahiro
Department of Reproductive and Developmental Biology, Institute of Medical Science, The University o
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KODAMA Tatsuhiko
Third Department of Internal Medicine, Faculty of Medicine, University of Tokyo
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Kodama Tatsuhiko
Third Department Of Internal Medicine Faculty Of Medicine University Of Tokyo
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UEDA Otoya
CSK Research Park Inc., Fuji Gotemba Research Laboratories, Chugai Pharmaceutical Co., Ltd.
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JISHAGE Kouichi
CSK Research Park Inc., Fuji Gotemba Research Laboratories, Chugai Pharmaceutical Co., Ltd.
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KAMADA Nobuo
CSK Research Park Inc., Fuji Gotemba Research Laboratories, Chugai Pharmaceutical Co., Ltd.
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SUZUKI Hiroshi
CSK Research Park Inc., Fuji Gotemba Research Laboratories, Chugai Pharmaceutical Co., Ltd.
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Kurihara Hiroki
Third Department Of Internal Medicine Faculty Of Medicine University Of Tokyo
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Toyoda Yutaka
Department Of Reproductive And Developmental Biology Institute Of Medical Science The University Of
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Suzuki H
National Research Center For Protozoan Diseases Obihiro University Of Agriculture And Veterinary Med
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Yazaki Yoshio
Third Department Of Internal Medicine Faculty Of Medicine The University Of Tokyo.
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AZUMA Sadahiro
Department of Animal Pathology, Institute of Medical Science,The University of Tokyo
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TOYODA Yutaka
Department of Animal Pathology, Institute of Medical Science,The University of Tokyo
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TOYODA Yutaka
Department of Reproductive and Developmental Biology, Institute of Medical Science, The University of Tokyo
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UEDA Otoya
CSK Research Park Inc.
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UEDA Otoya
CSK Research Park, INC.
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KAMADA Nobuo
CSK Research Park Inc.
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KAMADA Nobuo
CSK Research Park, INC.
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JISHAGE Kouichi
CSK Research Park, INC.
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Toyoda Yutaka
Department of Animal Pathology, Institute of Medical Science, University of Tokyo
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