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

将4mg/ml大鼠尾源性胶原（酸溶型-胶原）热变性100μl，在Transwell（Corstar，孔径0.3μm）中形成堵塞层。在这一层上以不同的细胞密度放置100μl的大鼠腹膜成纤维细胞分散胶原。用F12培养基孵育2 d后，按10^6/ml的细胞密度在Transwell膜的另一侧接种大鼠骨膜间皮细胞。在一周内形成间皮细胞层，呈多角形细胞形态，这在体内和体外标准细胞培养中都能观察到。在不同的胶原浓度和胶原层厚度下，测定尿素通过胶原层的通透性。据推测，尿素渗透性与胶原蛋白的厚度和浓度有关。这些结果表明，其他细胞外成分，如透明质酸，可能在溶质通过腹膜的运输中起关键作用，因为间皮下致密区增厚和透明质酸浓度增加通常在长期腹膜透析患者中发现。

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.