A Novel Construction Culture for Kidney Tissue and Evaluation for its Function in terms of Regeneration Therapy
| Project/Area Number |
15360434
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| Research Category |
Grant-in-Aid for Scientific Research (B)
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| Allocation Type | Single-year Grants |
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| Section | 一般 |
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| Research Field |
Biofunction/Bioprocess
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| Research Institution | University of Tsukuba |
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Principal Investigator |
WANG P-Chao University of Tsukuba, Graduate School of Life and Environmental Sciences, Professor, 大学院・生命環境科学研究科, 教授 (80261775)
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| Co-Investigator(Kenkyū-buntansha) |
SAITO Akira Tokai University, Institute of Medical Science, Professor, 総合医学研究所第三部門, 教授 (60307296)
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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 |
¥13,300,000 (Direct Cost: ¥13,300,000)
Fiscal Year 2005: ¥2,600,000 (Direct Cost: ¥2,600,000)
Fiscal Year 2004: ¥5,100,000 (Direct Cost: ¥5,100,000)
Fiscal Year 2003: ¥5,600,000 (Direct Cost: ¥5,600,000)
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| Keywords | renal glomeruli / epithelial cells / mesangial cells / metanephros / scaffold / collagen membrane / dialysis / tissue construction / TGFβ1 / 発生期後腎 / 尿管芽 / 腎糸宮体 / MMP / 毛細血管 / angiogenesis / vasculogenesis / 血管新生 / 間質系細胞 / 平面旋回培養 / disposable膜培養装置 / 擬微小重力 / 分化 / 振盪式膜培養装置 / 3次元細胞培養 / 微小重力 / 間質細胞 / transwell / 分枝 |
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| Research Abstract |
The concept of tissue engineering has been developed since early 1990. Three key factors including cell, cellular scaffold such as ECM (extra-cellular matrix) and cytokine such as growth factor are the basic materials used to form tissue. The successful example such as skin sheet has been available commercially. However, tissue comprising complicated structure and low regeneration ability such as kidney has not been constructed successfully. In this study, we focused on the regeneration of glomeruli, especially the glomerular epithelial cells for the chronic nephiritic disease. The symptom of epithelial cells of glomeruli for diabetes or Finnish disease is the foot process fusion which causes the disappearance of slit diaphragm between cells and impairs the polarity and flattens the morphology of epithelial cells.
Three techniques have been developed in this study. The first one is a technique combining the mesh sieving method and fine mince method to isolate glomeruli rapidly from mamm
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alian kidney and culture the epithelial cells of Bowman capsule, endothelial cells, mesangial cells and podocytes. The second one is to design a novel collagen membrane with high permeability supported by fibril gauge at the edge of collagen gel. The high permeability allows cytokines passing through membrane and the fibril gauge provides high absorption of nutrients and easy transport of the nutrients to the cells on the collagen gel. The third one is a new concept to co-culture various cells at the opposite sides of gauge-gel membrane. Using the high permeable gauge gel membrane, cellular paracrine and cell-cell interaction can be investigated. In this study, we co-cultured epithelial cells with endothelial cells and mesangial cells to see if the epithelial cell polarity can be regenerated in vitro. Generally, cells cultured on the dish show flattened shape and loose polarity. To mimicking the in vivo environment, collagen-gel-membrane is used as cellular scaffold, and epithelial cells and mesangial cells/endothelial cells were located at the opposite sides of scaffolds.
Our results showed that epithelial cells formed polarity only when co-culturing with mesangial cells. Further investigation revealed that mesangial cells in the co-cultured system increased its secretion of fibronectin which improved the focal contact of epithelial cells to the collagen gel-membrane and subsequently increased cell-cell interaction and therefore polarity formation was achieved. The collaborator of this research also carried out the in vivo transplantation to find the optimum conditions for epithelial cells culture. Slight improvement of the nephritic disease was confirmed.
However, epithelial cell is only one constitutive cell of glomeruli which cannot cover most diseases occurred to the various cells of glomeruli. This study provided us with a new direction to investigate the branching of uteretic bud, vasculogenisis and angiogenesis of metanephros which may lead us to find the key factors for a successful constitution of kidney in vitro. Less
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Report
(4 results)
Research Products
(37 results)
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[Journal Article] Screening for depression in hemodialysis patients : Assoications with diagnosis, treatment, and outcomes in the DOPPS2005
Author(s)
A.A.Lopes, J.M.Albert, E.W.Young, S.Satayathum, R.L.Pisoni, V.E.Andreucci, D.L.Mapes, N.A.Mason, S.Fukuhara, B.Wikstrom, A.Saito, F.K.Port
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Journal Title
Kidney Int. vol.66
Pages: 2047-2053
Description
「研究成果報告書概要(欧文)」より
Related Report
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