| Co-Investigator(Kenkyū-buntansha) |
FUJII Tohru Nagasaki University, Graduate School of Biomedical Sciences, Emeritus Professor, 大学院・医歯薬学総合研究科, 名誉教授 (60136661)
AKITA Sadonari Nagasaki University Medical and dental Hospital, Assistant Professor, 医学部・歯学部附属病院, 助手 (90315250)
HIRANO Akiyoshi Nagasaki University, Graduate School of Biomedical Sciences, Professor, 大学院・医歯薬学総合研究科, 教授 (90208835)
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| Research Abstract |
In order to investigate the receptor tyrosine kinase profiles, each component involving in the flap viability, such as epidermal keratinocyte, endothelial cell, dermal fibroblast and fetal dermal fibroblast was at first tested on the cell-to-cell interaction. The fetal fibroblast (BALB-3T3, clone A3) is well regulated signal-transduction pathway via leukemia inhibitory factor (LIE') associated receptor and intracellular mechanism such as STAT3 tyrosine phosphorylatiorn. The permanently transfected mouse full-length LIF cDNA into the ALB-3T3, clone A3 and the cell is passaged. The vascular endothelial growth facto (VEGF) significantly increased the cell growth in the doseincrease 1,10,100ng, dependent-manner. The phosphorylated-Erk p42/p44 Mitogen-Associated Protein (MAP), which is highly related to the proliferation of LIF-mediated cell, is induced from 1 to 30 minutes after 10ng VEGF and from l to 60 minutes after 100ng VEGF stimulation in the serum-free medium condition.
The modified
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Boyden dual chamber analyses were performed with interaction between the human mesenchymal stem cell (hMSC), which is a key role player of the ectodermal as well as mesenchymal lineage, and other, adult cell types such as epidermal keratinocyte, endothelial cell and dermal fibroblast. In 16-hour incubation with the hMSC in the top chamber and adult cells in the lower chamber demonstrated the significant cell migration in epidermal keratinocyte compared to endothelial cell and dermal fibroblast (345.0±61.50,36.2±14.45, 63.8±24.81, respectively, p<0.01). The monolayer co-culture of hMSC and epidermal keratinocyte demonstrated basal membrane-like protein production between the two cell types by ultrastructural analyses. The hMSC stimulated by basic fibroblast growth factor (bFGF) accelerated the skin defect together with artificial dermis template. These results may indicate the fetal fibroblast, which is gene-transfected of LIF cDNA, is useful in cell proliferation and further the mesenchymal stem cell may be a powerful tool for regenerative medicine. Less
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