de Groot C J, Chao V A, Roberts J M, Taylor R N
W. M. Keck Laboratory of Reproductive and Developmental Molecular Biology, Department of Obstetrics, Gynecology, and Reproductive Sciences, University of California School of Medicine, San Francisco 94143, USA.
Am J Physiol. 1995 Apr;268(4 Pt 2):H1613-20. doi: 10.1152/ajpheart.1995.268.4.H1613.
Human umbilical vein endothelial (HUVE) cells plated on plastic or gelatin-coated dishes grow as a "cobblestone" monolayer. By contrast, endothelial cells cultured on a complex matrix (e.g., Matrigel) form three-dimensional, capillary-like structures. In the current study, we verified the capillary phenotype of the latter structures and asked whether the morphological changes induced by extracellular matrix also affect human endothelial gene expression and function in vitro. Concentrations of cellular fibronectin, prostacyclin, and endothelin-1 were measured in the conditioned media by enzyme-linked immunosorbent and radioimmunoassays. Steady-state concentrations of HUVE mRNA were estimated by reverse transcription-polymerase chain reaction and quantified by Northern analyses to assess fibronectin and endothelin-1 gene expression. We found that the subjacent extracellular matrix affects the morphology, proliferation, and differentiation of HUVE cells in vitro. Cells cultured on gelatin were more mitotically active, expressed significantly less cellular fibronectin, made similar amounts of prostacyclin, and secreted significantly more endothelin-1 compared with the same cells grown on a Matrigel substrate.
接种在塑料或明胶包被培养皿上的人脐静脉内皮（HUVE）细胞呈“鹅卵石”样单层生长。相比之下，在复杂基质（如基质胶）上培养的内皮细胞形成三维的、毛细血管样结构。在本研究中，我们验证了后一种结构的毛细血管表型，并探讨细胞外基质诱导的形态变化是否也会影响人内皮细胞的基因表达和体外功能。通过酶联免疫吸附测定和放射免疫测定法测量条件培养基中细胞纤连蛋白、前列环素和内皮素-1的浓度。通过逆转录-聚合酶链反应估计HUVE mRNA的稳态浓度，并通过Northern分析进行定量，以评估纤连蛋白和内皮素-1基因的表达。我们发现，下层细胞外基质会影响HUVE细胞在体外的形态、增殖和分化。与在基质胶底物上生长的相同细胞相比，在明胶上培养的细胞有更强的有丝分裂活性，细胞纤连蛋白表达显著减少，前列环素产量相似，但内皮素-1分泌显著增加。
