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New cell culture method using type IV collagen

使用 IV 型胶原蛋白的新细胞培养方法

基本信息

批准号：
13558104
负责人：
IMAMURA Yasutada
金额：
$ 7.42万
依托单位：
The University of Tokyo
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
2001
资助国家：
日本
起止时间：
2001 至 2003
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-13558104/
关键词：
type IV collagen type IV collagen gel smooth muscle cell mesangium cell collagen gel basement membrane type V collagen メッシュワークコラーゲン会合体レンズカプセル線維芽細胞コラーゲンゲル多細胞メッシュワーク

项目摘要

This study aimed to explore and establish utilization of type IV collagen as culture substrate and culture method using the collagen. Transfer of some techniques related to manufacturing of the culture substrates was originally intended. Banned distribution of bovine derived material in Japan after first finding of a cattle affected with BSE made us to change our study plan. Since it was impossible to get the bovine sources, especially eye-related tissues such as lens capsules in Japan which is the best starting material for type IV collagen preparations, import of those from BSE-free country was successfully tried. But increased cost for the preparation resulted in small scale preparation only for laboratory use. New finding are summarized as follows :1.Improved recovery of type IV collagen from bovine lens capsuleWe obtained more efficient method with roughly twice amount to recover type IV collagen capable of forming gel. Type IV collagen in the preparation by the new method was par … More tially modified with some amino acid residues but still retained gel forming property on which human smooth muscle cells and human mesangium cells formed meshwork like cellular network.2.Trials of new cell culture methods using type IV collagena.A hat shape gelType IV collagen gel formed on a circle cover glass for cell culture support were getting dry from its periphery and remained in the center part of the circle. The shape of the substrate looks like a hat which is put on man's head and is called a hat shape gel. Same type IV collagens form two types of molecular aggregates between periphery and center. Smooth muscle cells cultured on the hat shape gel showed remarkable differences between both parts of the culture glass. The cells in the center part formed meshwork such as cells cultured on type IV collagen gel but those in periphery part did not form meshwork but grew well. The result demonstrated that physical property of the type IV collagen aggregates affects cellular response.b.Utilisation of type V collagen as cell culture substratesType V collagen distributes relatively close to basement membrane. Effect of type V collagen aggregates as culture substrates was examined and compared that of type IV collagen aggregates. Type V collagen aggregates affects unique effect on cellular response, such as shape and proliferation. Less

本研究旨在探索和建立IV型胶原蛋白作为培养基质的利用以及使用该胶原蛋白的培养方法。最初的目的是转让一些与培养基质制造相关的技术。首次发现感染疯牛病的牛后，日本禁止销售牛源材料，这促使我们改变了研究计划。由于在日本无法获得牛源，尤其是与眼睛相关的组织，例如晶状体囊，这是IV型胶原制剂的最佳原材料，因此成功尝试从无疯牛病国家进口。但制备成本的增加导致小规模制备仅供实验室使用。新发现概括如下： 1.提高从牛晶状体囊中回收IV型胶原蛋白我们获得了更有效的方法，大约用两倍的量回收能够形成凝胶的IV型胶原蛋白。新方法制备的IV型胶原蛋白被部分氨基酸残基修饰，但仍然保留了凝胶形成特性，人平滑肌细胞和人系膜细胞在其上形成网状细胞网络。2.使用IV型胶原蛋白的新细胞培养方法的试验。在用于细胞培养支持的圆形盖玻片上形成的帽子形状的凝胶IV型胶原蛋白凝胶从其外围变干并保持不变在圆的中心部分。基材的形状看起来像戴在人头上的帽子，称为帽子型凝胶。相同的 IV 型胶原蛋白在外围和中心之间形成两种类型的分子聚集体。在帽子形状凝胶上培养的平滑肌细胞在培养玻璃的两个部分之间显示出显着差异。中心部分的细胞形成网状结构，如在IV型胶原凝胶上培养的细胞，但外围部分的细胞没有形成网状结构，但生长良好。结果表明，IV型胶原聚集体的物理性质影响细胞反应。b.V型胶原作为细胞培养基质的利用V型胶原分布相对靠近基底膜。检查了 V 型胶原聚集体作为培养基质的效果，并比较了 IV 型胶原聚集体的效果。 V 型胶原蛋白聚集体对细胞反应（例如形状和增殖）有独特的影响。较少的