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Establishment of Novel Embryonic Stem Cell Lines Derived from the Common Marmoset (Callithrix jacchus)

^a Division of Laboratory Animal Science, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan;
^b Department of Urology, Urayasu Hospital, Juntendo University, Urayasu, Chiba, Japan;
^c Department of Molecular Genetics, Division of Molecular and Clinical Genetics, Medical Institute of Bioregulation, Kyushu University, Hakata, Fukuoka, Japan;
^d Tokai University School of Medicine, Isehara, Kanagawa, Japan;
^e Department of Physiology, Keio University School of Medicine, Shinjuku-ku, Tokyo, Japan;
^f Research Project Center,
^g Department of Genetics, Central Institute for Experimental Animals, Kawasaki, Kanagawa, Japan;
^h Division of Molecular Therapy, Institute of Medical Science, University of Tokyo,
ⁱ Institute of Obstetrics & Gynecology in Clinical Medicine, University of Tsukuba,
^j Laboratory of Embryonic Stem Cell Research, Stem Cell Research Center,
^k Department of Development and Differentiation, Institute for Frontier Medical Sciences, Kyoto University, Kyoto, Japan

Key Words. Embryonic stem cells • Common marmoset • Embryoid body • Nonhuman primate • Teratoma formation

Correspondence: Kenzaburo Tani, M.D., Ph.D., Department of Molecular Genetics, Division of Molecular and Clinical Genetics, Medical Institute of Bioregulation, Kyushu University, Hakata, Fukuoka 812-8582, Japan. Telephone: 81-92-642-6434; Fax: 81-92-642-6444; e-mail: taniken{at}bioreg.kyushu-u.ac.jp; for CMES cell distribution, contact Erika Sasaki at esasaki{at}ciea.or.jp

The successful establishment of human embryonic stem cell (hESC) lines has inaugurated a new era in regenerative medicine by facilitating the transplantation of differentiated ESCs to specific organs. However, problems with the safety and efficacy of hESC therapy in vivo remain to be resolved. Preclinical studies using animal model systems, including nonhuman primates, are essential to evaluate the safety and efficacy of hESC therapies. Previously, we demonstrated that common marmosets are suitable laboratory animal models for preclinical studies of hematopoietic stem cell therapies. As this animal model is also applicable to preclinical trials of ESC therapies, we have established novel common marmoset ESC (CMESC) lines. To obtain marmoset embryos, we developed a new embryo collection system, in which blastocysts can be obtained every 3 weeks from each marmoset pair. The inner cell mass was isolated by immunosurgery and plated on a mouse embryonic feeder layer. Some of the CMESC lines were cultured continuously for more than 1 year. These CMESC 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 CMESCs are pluripotent, as evidenced by in vivo teratoma formation in immunodeficient mice and in vitro differentiation experiments. Our established CMESC lines and the common marmoset provide an excellent experimental model system for understanding differentiation mechanisms, as well as the development of regenerative therapies using hESCs.