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

腹膜驻留细胞（人工腹膜）三维培养的研究

• 批准号: 10680810
• 负责人: HORIUCHI Takashi
• 金额: $ 1.92万
• 依托单位: University of East Asia
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10680810/
• 关键词: PERITONEAL DIALYSIS; PERITONEAL FIBROSIS; 3D CULTURE; SOLUTE TRANSPORT; 画像解析; 細胞遊走; 細胞増殖; 細胞培養; サイトカイン; RT-PCR; TGF-β1; 腹膜繊維芽細胞; RT-RCR; TGF-β

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.

通过热变性100μl 4 mg/ml大鼠尾衍生胶原（酸溶性型胶原），在Transwell（Corstar ; 0.3μm孔径）中形成栓塞层。在该层上，以不同的细胞密度放置100μl大鼠腹膜成纤维细胞分散的胶原。用F12培养基孵育2天后，将大鼠腹膜间皮细胞以10^6/ml的细胞密度接种在Transwell膜的另一侧。在一周内，形成了具有多边形细胞形态的间皮细胞层，这通常在体内和体外标准细胞培养中观察到。在不同的胶原蛋白浓度和层厚度下测量尿素通过胶原蛋白层的渗透性。据推测，尿素渗透性与胶原厚度及其浓度相关。这些结果表明，其他细胞外成分，如透明质酸可能有一个至关重要的作用，通过腹膜溶质运输，因为增厚的中皮下致密区和透明质酸浓度增加，通常发现在长期腹膜透析患者。

项目成果

Horiuchi T, Inoue T, et al: "Prolonged preservation of rat peritoneal mesothelial cell(RPMC)and rat peritoneal fibroblast(RPFB)by a glucose degradation product(GDP)depleted and neutral pH PD solution."Clin Exp Nephrol.. (submitted). (2001)

Horiuchi T、Inoue T 等人：“通过葡萄糖降解产物 (GDP) 耗尽和中性 pH PD 溶液延长大鼠腹膜间皮细胞 (RPMC) 和大鼠腹膜成纤维细胞 (RPFB) 的保存。”Clin Exp Nephrol..（已提交）

Sakai A,Horiuchi T,et al.: "Peritoneal Dialysis foltration(PDF)process with protein retaining dialysate for high-Rermeable patients" Pcritoneal Dialysis International. 19(S1). 9 (1999)

Sakai A、Horiuchi T 等人：“针对高可重复性患者的保留蛋白质透析液的腹膜透析滤过 (PDF) 过程”Pcritoneal Dialysis International。

Horiuchi, t: "kinetics in CAPD"Therapeutics & Engineering. 11(4). 630-633 (1999)

Horiuchi, t：“CAPD 动力学”

堀内 孝: "CAPDのKinetics"医工学治療. 11(4). 630-634 (1999)

Takashi Horiuchi：“CAPD 动力学”医学工程治疗 11(4) 630-634 (1999)。

大畑 淳,堀内 孝: "培養腹膜繊維芽細胞を用いた急性傷害モデルにおけるTGF-β1とVEGF産生遺伝子の発現"人工臓器. 29(3). S61 (2000)

Jun Ohata、Takashi Horiuchi：“使用培养的腹膜成纤维细胞在急性损伤模型中表达 TGF-β1 和 VEGF 生成基因”Artificial Organ S61 (2000)。