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Preparation of reconstituted extracellular matrices useful for cell culture

用于细胞培养的重组细胞外基质的制备

基本信息

批准号：
07558249
负责人：
HAYASHI Toshihiko
金额：
$ 2.18万
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 (A)
财政年份：
1995
资助国家：
日本
起止时间：
1995 至 1997
项目状态：
已结题

项目摘要

The objective of the present study is to elucidate the role of extracellular matrix, collagen protein family in particular, in maintaining the function of multicellular system through a synthetic methodology so that the system may show a novel function. Type I collagen gel culture containing fibroblasts within serves as a dermis-like replacement. We have obtained several bodies of unique information on the effects of type IV collagen gel and type V collagen fibrils on cellular behaviors in culture. The type IV collagen extracted from bovine lens capsule formed gel under unique conditions where specific interactions had been unexpected. Cells, hepatic stellate cells initially, cultured on the type IV collagen gel showed a number of unique behaviors that could never have been seen by the cultures on type I collagen gel or on type IV collagen without a gel form. The cell behaviors included extremely elongated cell morphology, formation of cell-to-cell junction through the tips of cell processes, and repressed proliferation. The observations are shared with other related cells such as smooth muscle cells and glomerular mesangium cells. Endothelial cells, epithelial cells or fibroblasts responded differently. Marker proteins for the contractile stage of smooth muscle cells were expressed when cultured on type IV collagen gel. Reconstituted type V collagen fibrils showed strong inhibition of growth of endothelial cells in spite of the strong cell adhesion.

本研究的目的是阐明细胞外基质，特别是胶原蛋白家族在维持多细胞系统功能中的作用，从而使多细胞系统显示出新的功能。含有成纤维细胞的I型胶原凝胶培养物可作为真皮样替代物。我们已经获得了一些关于IV型胶原凝胶和V型胶原纤维在培养中对细胞行为影响的独特信息。从牛晶状体囊膜中提取的IV型胶原在特殊的条件下形成凝胶，在这种条件下，特定的相互作用是意想不到的。最初在IV型胶原凝胶上培养的肝星状细胞表现出许多独特的行为，这些行为是在I型胶原凝胶上或在没有凝胶形式的IV型胶原上培养的。细胞行为包括极长的细胞形态，通过细胞突起尖端形成细胞与细胞的连接，并抑制增殖。这些观察结果与其他相关细胞，如平滑肌细胞和肾小球系膜细胞等也是相同的。内皮细胞、上皮细胞或成纤维细胞的反应不同。在IV型胶原凝胶上培养时，可表达平滑肌细胞收缩阶段的标志蛋白。重组V型胶原纤维虽然具有较强的细胞黏附性，但对内皮细胞的生长有较强的抑制作用。