STUDY ON THREE DIMENTIONAL CULTURE OF REPITONAL RESIDENT CELLS(AETIFICIAL PERITONEUM)

腹膜驻留细胞三维培养的研究

• 批准号: 11694181
• 负责人: HORIUCHI Takashi
• 金额: $ 6.14万
• 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 (B)
• 财政年份: 1999
• 资助国家: 日本
• 起止时间: 1999 至 2001
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-11694181/
• 关键词: 腹膜透析; 腹膜硬化症; 細胞培養; 三次元培養; 画像解析; 腹膜修復; 細胞増殖; 細胞遊走; サイトカイン; RT-PCR; VEGF; TGF-β1

Extracellular matrices layered with collagen and hyaluronan was formed in the Transwell (Corstar ; 0.3 μm pore size) and utilized for a solute-transfer experiment. Solutes with small molecular weight, except for urea, permeate these layers depending on concentration of both collagen and hyaluronan concentrations.In addition to solute-transfer experiments, a possible application of 3D model is a wound-healing study. To establish this experimental set-up, defoliation of mesothelial layer was made by a droplet of IN NaOH, and then microscopically observed by CCD camera. Digital images were aquired every 30 minutes until remesothelialization was completed. These date were then analysed to calculate cell size, rate of remesothelialization. 30〜4Q% increase of cell size were observed in comparison between pre and post healing. Digital video representation demonstrates cell migration prior to cell proliferation during wound healing on the monolayered culture of human peritoneal mesothelial cell.In conclusion 3D model of the peritoneum could be used for a various type of in vitro study such as solute-transfer experiment and wound healing experiment to elucidate fibrotic process of the peritoneum due to peritoneal dialysis procedure.

在Transwell（Corstar ; 0.3 μm孔径）中形成胶原和透明质酸分层的细胞外基质，并用于溶质转移实验。小分子量的溶质，除了尿素，渗透这些层取决于胶原蛋白和透明质酸的浓度concentrations.In除了溶质转移实验，3D模型的一个可能的应用是伤口愈合的研究。为了建立该实验装置，通过IN NaOH液滴进行间皮层的脱叶，然后通过CCD照相机进行显微观察。每30分钟采集一次数字图像，直到再间皮化完成。然后分析这些数据以计算细胞大小、再间皮化率。愈合前与愈合后相比，细胞大小增加了30 ~ 40%。数字视频演示了在单层培养的人腹膜间皮细胞的伤口愈合过程中细胞迁移之前的细胞增殖。结论腹膜的3D模型可用于各种类型的体外研究，如溶质转移实验和伤口愈合实验，以阐明腹膜透析过程中腹膜纤维化的过程。

项目成果

堀内孝: "腹膜硬化"CTPD. 19(3). 6-7 (2001)

Takashi Horiuchi：“腹膜硬化”CTPD 19(3) (2001)。

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

Horiuchi, t：“CAPD 动力学”

堀内 孝, その他: "培養細胞の障害修復過程の画像解析システムの開発"腎と透析. (2002)

Takashi Horiuchi 等人：“用于培养细胞修复过程的图像分析系统的开发”《肾脏与透析》（2002 年）。

Horiuchi T. et al.: "Expression of transforming growth factor (TGF-β1) mRNA and contractility of extracellular"Clin Exp Nephrol. 4. 105-113 (2000)

Horiuchi T.等：“转化生长因子(TGF-β1) mRNA的表达和细胞外的收缩性”Clin Exp Nephrol。 4. 105-113 (2000)

Horiuchi, T.Nagasawa, K. et al: "Effect of glucose degradation products (GDP) and pH in peritoneal dialysate on proliferation of rat peritoneal mesothelial cell and fibroblast"J Am Soc Nephrol. 11. 209A (2000)

Horiuchi, T.Nagasawa, K. 等人：“腹膜透析液中葡萄糖降解产物 (GDP) 和 pH 对大鼠腹膜间皮细胞和成纤维细胞增殖的影响”J Am Soc Nephrol。