| Co-Investigator(Kenkyū-buntansha) |
HATA Rhyichiro Kanagawa Dental College, School of Dentistry, Professor, 歯学部, 教授 (10014276)
ASASHIMA Makoto The University of Tokyo, Graduate School of Arts and Sciences, Professor, 総合文化研究科, 教授 (00090564)
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| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 2005: ¥900,000 (Direct Cost: ¥900,000)
Fiscal Year 2004: ¥1,300,000 (Direct Cost: ¥1,300,000)
Fiscal Year 2003: ¥1,300,000 (Direct Cost: ¥1,300,000)
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| Research Abstract |
The dissociated Xenopus animal cap cells treated with activin form an aggregate and it develops various tissues in vitro. In this study, to induce jaw cartilage from undifferentiated cells effectively, we developed a culture method to manipulate body patterning in vitro, using activin A and dissociated animal cap cells. An aggregate consisting only of activin A-treated dissociated cells developed into endodermal tissues. However, when activin A-treated cells were mixed with untreated cells at a ratio of 1:5,the aggregate developed cartilage with the maxillofacial regional marker genes, goosecoid, Xenopus Distal-less 4,and X-Hoxa2. When this aggregate was transplanted into the abdominal region of host embryos, maxillofacial structures containing cartilage and eye developed. We raised these embryos to adulthood, and found that tooth germ had developed in the transplanted tissue. Furthermore, we have developed a chemically defined serum-free medium, designated ESF7,in which leukemia inhib
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itory factor(LIF) clearly stimulated murine ES cell proliferation accompanied by increased expression of nanog and Rex-1 and decreased FGF-5 expression. These effects were dependent upon the concentration of LIF. In ESF7 medium, ES cells maintained an undifferentiated phenotype as manifested by the expression of the transcription factor Oct3/4,the stem cell marker SSEA-1,and alkaline phosphatase. Withdrawal of LIF from ESF7 medium resulted in ES cell apoptosis. Addition of serum to ESF7 medium promoted ES cell differentiation. Addition of BMP4 promoted ES cell differentiation into simple epithelial-like cells. By contrast, FGF-2 promoted ES cell differentiation into neuronal and glial-like cells. As this simple serum-free adherent monoculture system supports the long-term propagation of pluripotent ES cells in vitro, it will allow the elucidation of ES cell responses to growth factors under defined conditions. Using these experimental system, we believe that we could develop the application system for the induction of tooth and jaws in future. Less
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