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a Division of Laboratory Animal Research, Research Center for Human and Environmental Sciences, Shinshu University;
b Department of Surgery, Shinshu University School of Medicine;
c Department of Pathology, Shinshu University School of Medicine, Shinshu, Japan
Key Words. Embryonic stem cell • Liver • Organogenesis • Cardiomyocyte • Endothelial cell • In vitro system
Correspondence: Yoh-ichi Tagawa, Ph.D., Department of Biomolecular Engineering, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, B-51 4259 Nagatsuta-cho, Midori-ku, Yokohama, Kanagawa, 226-8501 Japan. Telephone: 81-45-924-5791; Fax: 81-45-924-5815; e-mail: ytagawa{at}bio.titech.ac.jp
Recent studies in the field of regenerative medicine have exploited the pluripotency of embryonic stem (ES) cells to generate a variety of cell lineages. However, the target has always been only a single lineage, which was isolated from other differentiated cell populations. In the present study, we selected sublines with a high capability for differentiation to contracting cardiomyocytes and also produced germ-line chimeric mice from a parent ES line. We also succeed in establishing embryoid bodies prepared from the ES cells that differentiated into not only hepatocytes but also at least two mesodermal lineages: cardiomyocytes that supported liver development and endothelial cells corresponding to sinusoids. This allowed the development of an in vitro system using murine ES cells that approximated the events of liver development in vivo. The expression of albumin was significantly higher in cardiomyocytes that had arisen in differentiated ES cells than in those that had not. Our in vitro system for liver organogenesis consists of a blood/sinusoid vascular-like network and hepatocyte layers and shows higher levels of hepatic function, such as albumin production and ammonia degradation, than hepatic cell lines and primary cultures of murine adult hepatocytes. This innovative system will lead to the development of second-generation regenerative medicine techniques using ES cells and is expected to be useful for the development of bioartificial liver systems and drug-metabolism assays.
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