A Method for the Selection of Human Embryonic Stem Cell Sublines with High Replating Efficiency After Single-Cell Dissociation

Authors

  • Kouichi Hasegawa,

    1. Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Kyoto University, Kyoto, Japan
    2. Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan
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  • Tsuyoshi Fujioka,

    1. Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Kyoto University, Kyoto, Japan
    2. Cell Engineering Division, BioResource Center, RIKEN, Tsukuba, Ibaraki, Japan
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  • Yukio Nakamura,

    1. Cell Engineering Division, BioResource Center, RIKEN, Tsukuba, Ibaraki, Japan
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  • Norio Nakatsuji,

    1. Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan
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  • Hirofumi Suemori Ph.D.

    Corresponding author
    1. Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Kyoto University, Kyoto, Japan
    • Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical Science, Kyoto University, 53 Kawaharacho, Shogoin, Sakyo-ku, Kyoto, 606-8507, Japan. Telephone: 81-75-751-3821; Fax: 81-75-751-3890
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Abstract

Human embryonic stem cells (hESCs) exhibit pluripotency and indefinite proliferation and are a potential source of cells for transplantation therapies and drug discovery. These applications will require large amounts of hESCs. However, hESCs are difficult to culture and maintain at larger scales, in part because of their low resistance to dissociation during passaging. To circumvent this, we developed a simple and easy method for establishing hESC sublines tolerant of complete dissociation. These cells exhibit high replating efficiency and also high cloning efficiency, and they maintain their ability to differentiate into the three germ layers. Several sublines have no detectable abnormalities in their karyotypes, and they retained their characteristics under feeder-free culture conditions and after freeze-thawing. Thus, these hESC sublines would be valuable for hESC applications.

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