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Original Article |
1 Division of Laboratory Animal Science, Central Institute for Experimental Animals, Japan
2 Department of Urology, Urayasu Hospital, Juntendo University, Japan
3 Department of Molecular Genetics, Division of Molecular and Clinical Genetics, Medical Institute of Bioregulation, Kyushu University, Japan
4 Tokai University School of Medicine, Japan
5 Department of Physiology, Keio University School of Medicine, Japan
6 Research Project Center, Central Institute for Experimental Animals, Japan
7 Department of Genetics, Central Institute for Experimental Animals, Japan
8 Division of Molecular Therapy, Institute of Medical Science, University of Tokyo
9 Institute of Obstetrics and Gynecology in Clinical Medicine, University of Tsukuba
10 Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center, Institute for Frontier Medical Sciences, Kyoto University
11 Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University
12 Division of Molecular and Clinical Genetics, Medical Institute of Bioregulation, Kyushu University, Japan
* To whom correspondence should be addressed. E-mail: taniken{at}bioreg.kyushu-u.ac.jp.
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Abstract |
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The successful establishment of human embryonic stem (ES) cell lines has inaugurated a new era in regenerative medicine by facilitating the transplantation of differentiated embryonic stem cells to specific organs. However, problems with the safety and efficacy of human embryonic stem cell therapy in vivo remain to be resolved. Pre-clinical studies using animal model systems, including non-human primates, are essential to evaluate the safety and efficacy of human embryonic stem cell therapies. Previously, we demonstrated that common marmosets are suitable laboratory animal models for pre-clinical studies of hematopoietic stem cell therapies. As this animal model is also applicable to pre-clinical trials of ES cell therapies, we have established novel common marmoset embryonic stem (CMES) cell lines. To obtain marmoset embryos, we developed a new embryo collection system, in which blastocysts can be obtained every three weeks from each marmoset pair. The inner cell mass (ICM) was isolated by immunosurgery and plated on a mouse embryonic feeder (MEF) layer. Some of the CMES lines were cultured continuously for more than one year. These CMES cell lines showed alkaline phosphatase activity, and expressed stage-specific embryonic antigen (SSEA)-3, SSEA-4, TRA-1-60, and TRA-1-81. On the other hand, SSEA-1 was not detected. Furthermore, our novel CMES cells are pluripotent, as evidenced by in vivo teratoma formation in immunodeficient mice and in vitro differentiation experiments. Our established CMES cell lines and the common marmoset provide an excellent experimental model system for understanding differentiation mechanisms, as well for the development of regenerative therapies using human ES cells.
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