Kay E P, Smith R E, Nimni M E
J Biol Chem. 1985 Apr 25;260(8):5139-46.
When primary corneal endothelial cells were grown in polymorphonuclear leukocyte (PMN)-conditioned medium, a minor population of cells acquired fibroblastic morphology. Such modulated endothelial cells supported by PMN-conditioned medium grew much faster than the major nonresponding polygonal endothelial cell. Upon serial passages, the modulated endothelial cells became the dominant cell type and eventually formed a homogeneous fibroblastic culture. At the same time, phenotypic changes of collagen were observed. The primary endothelial cells grown in PMN-conditioned medium, consisting of responding elongated cells and nonresponding polygonal endothelial cells, produced predominantly type IV collagen with type III collagen as a minor component. As cells were subcultured and fibroblastic cells became dominant, type IV collagen synthesis was dramatically decreased and type I collagen synthesis was increased in parallel. When they reached the fully modulated stage, the cultures synthesized types I and III collagen, with type I accounting for 75-85% of the total. Type I collagen synthesized by the fibroblastic endothelial cells shared common characteristics with known type I collagen, such as migration behavior on sodium dodecyl sulfate-polyacrylamide gel electrophoresis, CNBr peptide profiles, and immunologic identity. Thus, PMNs apparently contribute to the modulation of corneal endothelial cells, causing them to acquire characteristics of fibroblasts, cell multilayering, and deposition of interstitial extracellular matrix composed predominantly of interstitial type I collagen.
当原代角膜内皮细胞在多形核白细胞(PMN)条件培养基中培养时,一小部分细胞获得了成纤维细胞形态。由PMN条件培养基支持的这种调节后的内皮细胞比主要无反应的多边形内皮细胞生长得快得多。经过连续传代,调节后的内皮细胞成为主要细胞类型,并最终形成均匀的成纤维细胞培养物。与此同时,观察到了胶原蛋白的表型变化。在PMN条件培养基中生长的原代内皮细胞,由有反应的细长细胞和无反应的多边形内皮细胞组成,主要产生IV型胶原蛋白,III型胶原蛋白为次要成分。随着细胞传代培养且成纤维细胞占主导,IV型胶原蛋白合成显著减少,I型胶原蛋白合成相应增加。当它们达到完全调节阶段时,培养物合成I型和III型胶原蛋白,其中I型占总量的75 - 85%。成纤维细胞样内皮细胞合成的I型胶原蛋白与已知的I型胶原蛋白具有共同特征,如在十二烷基硫酸钠 - 聚丙烯酰胺凝胶电泳上的迁移行为、溴化氰肽谱和免疫特性。因此,PMN显然有助于角膜内皮细胞的调节,使其获得成纤维细胞的特征、细胞多层化以及主要由间质I型胶原蛋白组成的间质细胞外基质的沉积。