STUDY ON THREE DIMENTIONAL CULTURE OF PERITONEAL RESIDENT CELLS(ARTIFICIAL PERITONEUM)

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Plug layer was formed in the Transwell(Corstar ; 0.3μm pore size)by thermally denaturing 100μl of 4mg/ml rat-tail derived collagen(acid soluble type- collagen). On this layer 100μl of rat peritoneal fibroblast dispersed collagen was placed at varying cell density. After incubating with F12 culture medium for 2 days, rat periotoneal mesothelial cell was seeded at cell density of 10^6/ml on the opposite side of the Transwell membrane. Within one week mesothelial cell layer was formed with polygonal cell morphology which is typically observed in vivo and in vitro standard cell culture. Permeability of urea though the collagen layer was measured at varying concentration of collagen and thickness of the layer. As is presumed, urea permeability was correlated to thichness of collagen and its concentration. These results suggest that the other extra-cellular component such as hyaluronan could have a crucial role in solute transport via the peritoneum because thickening of the submesothelial compact zone and increased hyaluronan concentration were typically found in the patients with a long-term peritoneal dialysis.