Three-dimensional multilayers of smooth muscle cells as a new experimental model for vascular biology

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Elastic fiber formation is disrupted with age and by health conditions including aneurysms and atherosclerosis. Despite considerable progress in the understanding of elastogenesis using the planar culture system and genetically modified animals, it remains difficult to restore elastic fibers in diseased vessels. To further study the molecular mechanisms, we created in vitro three-dimensional vascular constructs. Three-dimensional cellular multilayers (3DCMs), which consisted of seven layers of neonatal rat aortic SMCs cultured in 1% fetal bovine serum in DMEM medium, exhibited layered elastic fibers and differentiated smooth muscle cell phenotype within seven days of being in a static culture condition. Furthermore, infiltration of THP-1-derived macrophages decreased the surrounding elastic fiber formation in 3DCMs. 3DCMs may offer a new experimental vascular model to explore pharmacological therapeutic strategies for abnormal vascular remodeling.