Human umbilical vein endothelial cells behavior on fibroblast-derived matrix via a physical or chemical modification

Human umbilical vein endothelial cells behavior on fibroblast-derived matrix via a physical or chemical modification
두 핑라메쉬수하에리박귀덕
Fibroblast-derived matrix (FDM); human umbilical vein endothelial cells; vascular morphogenesis; matrix stiffness; matrix remodeling
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Extracellular matrix (ECM) macromolecules are closely involved in the process of blood vessel formation, remodeling, and regression. In this study, fibroblast-derived matrix (FDM) was used to investigate the impact of two-dimensional (2D) ECM microenvironment on vascular morphogenesis of endothelial cells (ECs) in vitro. FDM was prepared by decellularizing in vitro-cultured NIH3T3 fibroblasts on the coverslips. These FDMs were then physically modified via different storage conditions (fresh, frozen, and air-dried). Physically modified FDMs exhibited a unique surface morphology and topological structure. Fresh FDM was the best in promoting ECs proliferation and forming an EC-specific capillary-like structure (CLS). It was notable that a time-dependent matrix remodeling by ECs was observed only in fresh FDM as illustrated by immunofluorescence staining against ECM macromolecules, such as laminin and fibronectin. Furthermore, cell migration was also more active on fresh FDM compared with frozen or dried FDMs as observed by live imaging microscope. Meanwhile, FDMs were subjected to a chemical modification using a crosslinking agent. Crosslinked FDM exhibited an increased stiffness up to 10KPa with chemical concentrations as determined via Nanowizard II bio-atomic force microscope. Both CLS formation and matrix remodeling are much more competitive with non-crosslinked, fresh FDM compared to the crosslinked ones, suggesting that ECs are very sensitive to matrix stiffness. Further analysis of microarray revealed some important regulatory factors on matrix remodeling and angiogenesis. Taken together, FDM provides a very effective model in the study of vascular morphogenesis of ECs on 2D platform in vitro.
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