| Budget Amount *help |
¥3,500,000 (Direct Cost: ¥3,500,000)
Fiscal Year 1999: ¥1,700,000 (Direct Cost: ¥1,700,000)
Fiscal Year 1998: ¥1,800,000 (Direct Cost: ¥1,800,000)
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| Research Abstract |
The objective of this study is to construct artificial kidney unit by means of (1) manupulating genes to enhance the cell capability to remove immune complex and induce the ability of immune tolerance, (2) design of the in vitro environment for cell culture using basement membrane-like matrix, (3) explant of the functional cells on the designed matrix, and finally to combine the tissue engineering technique to achieve a convenient, functional artificial kidney.
Previously, we have reported that the gene (CR1) related to the immune complex removal and gene (CTLA4-Ig) related to the immune tolerance have been transfected to glomerular epithelial cells, and the transfection of genes pE6 and pE7 to glomerular mesangial cells had successfully immortalized cells.
This year, we investigated the effect of cll-cell interaction between glomerular epithelial cell and mesangial cell and found the optimum conditions for coculture of cells. Moreover, excellular matrix (ECM and collagen gel) for cell c
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ulture were also investigated. Furthermore, we constructed a naturally branched gauge gel by immersing gauge in collagen gel added with ascorbic acid and followed by UV radiation. The above two kinds of cells were explanted to each side of the formed gel.
The results showed that ECM caused glomerular epithelial cell to form honey-cobbled shape and depress the contractility of mesangial cell. On the other hand, collagen gel caused epithelial cells to aggregate, but addition of ascorbic acid showed a smooth outgrowth for both cells. Coculture of both cells on the gauge gel resulted in a well growth of both cells where the epithelical cell showed a sheet-like form quickly, and mesangial cells adhered to the branch of gauge and aggregated gradually. FDA and PI demonstrated that both cells were viable in the coculture for more than 1 week without medium change. This fact may imply that the gauge has tranproted medium nutrient and oxygen to cells and can be used as a soft scaffold for the coculture of cells. Less
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