Development of Cultured Renal Tubules with Cell Sheet Engineering

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We developed a temperature-responsive cultured dishes by covalent grafting of poly (N-isopropylacrylamide) which shows the hydrophilic/hydrophobic change of surface property. This surface shows weak hydrophobicity similarly to commercial tissue culture polystyrene dishes and allows various cell adhesion and spreading on it at 37℃. But the cells are detached from the highly hydrated surfaces without any need of proteolytic enzymes such as trypsin after reducing temperature below 32℃. All the spread cells connecting via cell-cell junctions are harvested as a single contiguous cell sheet. In the present study, we examined the utilization of harvested kidney epithelial cell sheets in the development of bioartificail renal tubules by combining the cell sheets and porous membranes for dialysis in terms of molecular transport and production. We developed a non-invasive cell sheet manipulation method from culture dishes to other surfaces such as porous membranes. Experimental devices were prepared by combining of primary human kidney epithelial cells and porous membranes. The expression and localization of various cell surface proteins are confirmed by immunostaining and confocal laser scanning microscopy.