Abstract
The purpose of this study was to test the feasibility of applying a novel coculture model for the investigation of endothelium-derived relaxing factor (EDRF) as well as exploring its applicability for investigating important cell-to-cell interactions. Bovine aortic endothelial cells (EC) were grown on micropore filters while porcine aortic smooth muscle cells (SMC) were grown separately on plates. When confluent these two cell layers were cocultured such that the EC maintained proper polarity and orientation to the underlying SMC. We found that coculturing EC with SMC for five minutes caused significant increase in SMC cGMP, 43±4 vs 268±13 fmols/well (p<0.0001). This EC-mediated effect was further augmented with EDRF agonists and with L-arginine supplementation, but was inhibited by nitro-L-arginine methyl ester (NAME) and reduced hemoglobin. When the EC were cocultured with subendothelial THP-1 monocytes for three hours prior to the SMC coculture, the EC mediated increase in SMC cGMP, both stimulated and unstimulated, was significantly reduced. We conclude that this flexible coculture model can be used to study EDRF release from EC and can be applied to study important cell-to-cell interactions that have been difficult to address in other models.
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Oskarsson, H., Hofmeyer, T. & Stoll, L.L. Demonstration of endothelium-mediated increase in vascular smooth muscle cGMP, in a flexible coculture model. Methods Cell Sci 17, 237–244 (1995). https://doi.org/10.1007/BF00986228
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DOI: https://doi.org/10.1007/BF00986228