Kennedy A, Frank R N, Sotolongo L B, Das A, Zhang N L
Kresge Eye Institute, Wayne State University School of Medicine, Detroit, MI 48201.
Curr Eye Res. 1990 Apr;9(4):307-22. doi: 10.3109/02713689008999619.
To investigate the effects of extracellular matrix components on cellular function, we cultured several types of ocular cells on substrates composed of extracellular matrix materials that were layered on culture dishes either as dried films or as gels. We measured cellular proliferation on these substrates and on a series of gels composed of varying proportions of rat tail tendon type I collagen and Matrigel, a commercially available extract of a basement membrane-producing murine tumor. In addition, we studied the biosynthesis of collagens and of proteoglycans by these cultured cells using [3H]-L-proline and [35S]-sulfate. The proliferative abilities of the various types of ocular cells on the dried film substrates, on uncoated plastic culture vessels, and on pure type I collagen gel, were similar. However, proliferation of ocular cells cultured on gels composed of greater than or equal to 90% Matrigel was markedly reduced. There was little or no inhibition of growth of two types of non-ocular cells: rat C6 astrocytoma cells, and human dermal fibroblasts. Histologic studies showed that the ocular cells tested often formed long strands and capillary-like tubes, and tended to "burrow" beneath the surface of substrates containing high percentages of Matrigel. Fibroblasts infrequently formed tubes, and exhibited the burrowing property also on gels containing primarily type I collagen, while C6 cells showed neither of these behaviors on any of the matrices tested. The elution pattern of newly synthesized [3H]-labeled and [35S]-labeled macromolecules produced by all of the cultured cell types, and detected by Sepharose CL-4B chromatography in the medium and in the cell layer plus matrix fractions did not vary following culture on the different substrates. Approximately twofold more of the newly synthesized collagens and proteoglycans were deposited in the cell layer plus matrix, and proportionately less appeared in the medium, when cells were cultured on type I collagen gels and on Matrigel than on the dried film substrates. These experiments demonstrate the influence of the extracellular matrix on several aspects of cell behavior, and provide further evidence that modification of the composition of the extracellular matrix may be an important determinant of normal or pathological cell function.
为了研究细胞外基质成分对细胞功能的影响，我们将几种类型的眼细胞培养在由细胞外基质材料构成的底物上，这些材料以干膜或凝胶的形式铺在培养皿上。我们测量了这些底物以及一系列由不同比例的大鼠尾腱I型胶原和基质胶（一种可商购的产生基底膜的小鼠肿瘤提取物）组成的凝胶上的细胞增殖情况。此外，我们使用[3H]-L-脯氨酸和[35S]-硫酸盐研究了这些培养细胞中胶原蛋白和蛋白聚糖的生物合成。各种类型的眼细胞在干膜底物、未包被的塑料培养容器以及纯I型胶原凝胶上的增殖能力相似。然而，在由大于或等于90%的基质胶组成的凝胶上培养的眼细胞增殖明显减少。两种非眼细胞：大鼠C6星形细胞瘤细胞和人皮肤成纤维细胞的生长几乎没有受到抑制。组织学研究表明，所测试的眼细胞经常形成长链和毛细血管样管，并倾向于在含有高百分比基质胶的底物表面下“钻入”。成纤维细胞很少形成管，并且在主要含有I型胶原的凝胶上也表现出钻入特性，而C6细胞在任何测试的基质上都没有表现出这两种行为。通过Sepharose CL-4B色谱法在培养基以及细胞层加基质部分中检测到的所有培养细胞类型产生的新合成的[3H]标记和[35S]标记的大分子的洗脱模式，在不同底物上培养后没有变化。当细胞在I型胶原凝胶和基质胶上培养时，与在干膜底物上培养相比，新合成的胶原蛋白和蛋白聚糖在细胞层加基质中的沉积大约多两倍，而在培养基中的出现比例相应减少。这些实验证明了细胞外基质对细胞行为多个方面的影响，并提供了进一步的证据表明细胞外基质组成的改变可能是正常或病理细胞功能的重要决定因素。
