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Embryonic Stem Cells |
1 Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan; Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan
2 Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Kyoto University, Kyoto, Japan; Cell Engineering Division, BioResource Center, RIKEN, Ibaraki, Japan
3 Cell Engineering Division, BioResource Center, RIKEN, Ibaraki, Japan
4 Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan
5 Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan
* To whom correspondence should be addressed. E-mail: hsuemori{at}frontier.kyoto-u.ac.jp.
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Abstract |
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Human embryonic stem (hES) cells 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 hES cells. However, hES cells 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 hES cell sub-lines 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 sub-lines have no detectable abnormalities in their karyotypes, and they retained their characteristics under feeder-free culture conditions and after freeze-thawing. Thus, these hES cell sub-lines would be valuable for hES cell applications.
Key Words. Human embryonic stem cells, replating efficiency, cloning efficiency
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