| Project/Area Number |
15590179
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| Research Category |
Grant-in-Aid for Scientific Research (C)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
General anatomy (including Histology/Embryology)
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| Research Institution | University of Occupational and Environmental Health |
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Principal Investigator |
DOI Yoshiaki University of Occupational and Environmental Health, School of Medicine, Department of Anatomy, Professor, 医学部, 教授 (30258602)
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| Co-Investigator(Kenkyū-buntansha) |
KUDO Hideaki University of Occupational and Environmental Health, School of Medicine, Department of Anatomy, Assistant Professor, 医学部, 講師 (40289575)
FUJIMOTO Sunao Nakamura Gakuen University Graduate School, Health and Nutritional Sciences, Professor, 大学院, 教授 (80080547)
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| Project Period (FY) |
2003 – 2005
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| Project Status |
Completed (Fiscal Year 2005)
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| Budget Amount *help |
¥2,200,000 (Direct Cost: ¥2,200,000)
Fiscal Year 2005: ¥300,000 (Direct Cost: ¥300,000)
Fiscal Year 2004: ¥700,000 (Direct Cost: ¥700,000)
Fiscal Year 2003: ¥1,200,000 (Direct Cost: ¥1,200,000)
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| Keywords | vasculogenesis / wound healing / endothelial cell / endothelial progenitor cell / VEGF / basic FGF / fibroblast / cell culture / 熱傷モデル / BrdU |
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| Research Abstract |
Vasculogenesis is defined as a neovascularization process by transformation of vasoformative mesenchymal cells to endothelial progenitor cells (EPCs). Although recent researchers insisted that bone marrow-derived EPCs via peripheral blood vessels are mainly involved in vasculogenesis during wound healing process of adult tissues, we have reported that multipotent mesenchymal cells derived from tissue fibroblasts located in several adult tissues transform to EPCs and participate in the acceleration of the vascular growth. This study deals with ultrastructural and immunocytochemical analyses on EPCs derived from cultured fibroblasts of the rat dermis. Deeply anesthetized adult Wistar rats received skin-biopsy in their back skin, and then fibroblasts were cultured on collagen-coated cell dishes. These cells were maintained in a humidified incubator containing a gas mixure of 5% CO_2 in 95% air at 37℃, and fed a cultured medium supplemented with fetal bovine serum, 10ng/ml basic fibroblast
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growth factor and 10ng/ml Vascular endothelial groeth factor. These cultured cells were prepared for light microscopic immunocytochemistry and electron microscopy, respectively. These cells, which reached monolayer confluence in the first time, showed no immunoreactivities for von Willebrand factor. After several days, some cultured cells possessed an abundance of cytoplasmic projections and were occasionally in contact with the neighboring ones forming solid cell cords. Since these cells showed immunoreactivities for von Willebrand factor and possessed many Weibel-Palade (WP) bodies, we defined them as phenotypes of EPCs. In addition, immunoreactivities for endothelin (ET)-1, big ET-1 and ET converting enzayme (ECE)-1 were seen in such EPCs. Since it is widely accepted that ET-1 and ECE-1 are stored in WP bodies of endothelial cells, we consider now that fibroblast-derived EPCs which are characterized by possessing WP bodies may be already transformed into endothelial cells and have ability to synthesize ET-1. Less
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