Endothelial cell adhesion, signaling, and morphogenesis in fibroblast-derived matrix.
Endothelial cell adhesion, signaling, and morphogenesis in fibroblast-derived matrix.
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DOI：
10.1016/j.matbio.2009.04.005
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发表时间：
2009-06
期刊：
影响因子：
--
通讯作者：
Patricia A. Soucy;L. Romer
中科院分区：
文献类型：
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作者：
Patricia A. Soucy;L. Romer
细胞外基质通过提供指导形态发生的信号线索在细胞发育中起着关键作用。为了研究天然基质提供的线索和内皮细胞对该信息的反应，使用人胎儿肺成纤维细胞来产生纤维三维基质。通过去污剂提取去除成纤维细胞后，通过免疫荧光和免疫印迹法鉴定剩余基质的蛋白质和蛋白聚糖成分。基质成分包括纤连蛋白、腱生蛋白-C、胶原蛋白I、胶原蛋白IV、胶原蛋白VI、多功能蛋白聚糖和核心蛋白聚糖。共定位分析表明，纤维连接蛋白是一种独特分布的基质蛋白。形态学，三维基质粘附，和整合素介导的信号在血管生成过程中，然后在人内皮细胞接种到成纤维细胞衍生的矩阵进行了研究。与纤连蛋白包被的盖玻片上的细胞相比，观察到细胞形态延长和细胞面积减少。细胞-基质粘附包含黏着斑蛋白、pY 397-FAK和pY 410-p130 Cas，并且所有这些与纤连蛋白的共定位多于与生腱蛋白-C、胶原蛋白I或胶原蛋白VI的共定位。此外，内皮细胞重塑成纤维细胞衍生的基质，并形成具有明显管腔的管网络。这些管中的基质粘附也主要与纤连蛋白共定位。1型基质金属蛋白酶在内皮细胞中的表达模式表明其参与了肾小管形成过程中发生的基质重塑。这些结果表明成纤维细胞来源的基质中的信息促进血管生成行为。
Extracellular matrix plays a critical role in cellular development by providing signaling cues that direct morphogenesis. In order to study both the cues that natural matrix provides and endothelial cell responses to that information, human fetal lung fibroblasts were used to produce a fibrous three-dimensional matrix. Following the removal of the fibroblasts by detergent extraction, protein and proteoglycan constituents of the remaining matrix were identified by immunofluorescence and immunoblotting. Matrix components included fibronectin, tenascin-C, collagen I, collagen IV, collagen VI, versican, and decorin. Colocalization analysis suggested that fibronectin was a uniquely distributed matrix protein. Morphology, three-dimensional matrix adhesions, and integrin-mediated signaling during vasculogenesis were then studied in human endothelial cells seeded onto the fibroblast-derived matrix. Elongated morphology and decreased cell area were noted, as compared with cells on fibronectin-coated coverslips. Cell-matrix adhesions contained vinculin, pY397-FAK, and pY410-p130Cas, and all of these colocalized more with fibronectin than tenascin-C, collagen I, or collagen VI. Additionally, the endothelial cells remodeled the fibroblast-derived matrix and formed networks of tubes with demonstrable lumens. Matrix adhesions in these tubes also predominantly colocalized with fibronectin. The pattern of membrane type 1 matrix metalloprotease expression in the endothelial cells suggested its involvement in the matrix remodeling that occurred during tubulogenesis. These results indicated that information in fibroblast-derived matrix promoted vasculogenic behavior.