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KMID : 1094720000050020092
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Biotechnology and Bioprocess Engineering
2000 Volume.5 No. 2 p.92 ~ p.98
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Immortalization of rat kidney glomerular mesangial cell and its coculture with glomerular epithelial cell
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Kida Toshinobu
Fujishima Sachi
Matsumura Masatoshi
Wang Pi-Chao
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Abstract
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Mesangial cell has several key roles in the control of glomerular function: it participates in the regulation of glomerular filtration rate, macromolecular clearance, and as both a source and target of numerous hormones and autocrines. Many of these insights into mesangial cell function have been obtained by studying mesangial cells in culture. However, no suitable cell lines have been established yet. We here reported the immortalization of rat kidney glomerular mesangial cell by transfection of E6 and E7 genes of human papillomavirus type 16 (HPV-16) via electroporation and lipofection. The results showed that only electroporation could transfect the genes to mesangial cells and the transfected cells maintained the viability for longer than 6 months. Fluorescence microscopic observation showed that cellular contractility and phagocytosis, which are the two main phenotypes of mesangial cells, are well maintained after transfection. The coculture of transfected mesangial cells with rat glomerular epithelial cells showed that the growth of mesangial cells was suppressed by epithelial cell, but the growth of epithelial cells was enhanced by mesangial cells. Moreover, an enhancing effect on the phagocytosis of mesangial cell was also observed in coculture. Such results may imply that the glomerular cell-cell interaction plays an important role in the regulation of cell proliferation and differentiation.
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KEYWORD
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immortalization, glomerular mesangial cell, glomerular epithelial cell, E6 and E7 genes, cellular contractility, phagocytosis
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