Preparation of reconstituted extracellular matrices useful for cell culture

本研究的目的是通过合成方法阐明细胞外基质，特别是胶原蛋白家族在维持多细胞系统功能中的作用，以便该系统可能显示出新的功能。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型胶原原纤维对培养细胞行为的影响的独特信息。从牛晶状体囊中提取的IV型胶原蛋白在意想不到的特定相互作用的特殊条件下形成凝胶。细胞，最初的肝星状细胞，在IV型胶原凝胶上培养，显示出许多独特的行为，这些行为在I型胶原凝胶和IV型胶原凝胶上的培养中是看不到的。细胞行为包括细胞形态极度拉长，细胞突起尖端形成细胞间连接，增殖受到抑制。其他相关细胞如平滑肌细胞和肾小球系膜细胞也有相同的观察结果。内皮细胞、上皮细胞和成纤维细胞的反应不同。在IV型胶原凝胶上培养平滑肌细胞，表达平滑肌细胞收缩期的标志蛋白。重组V型胶原原纤维虽具有较强的细胞粘附性，但对内皮细胞的生长有较强的抑制作用。

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.