Development of a 3- dimensional culture model system simulating arterial intima and its application for atherosclerosis study.

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1. Development of a 3-dimensional (3-D) culture system simulating arterial intimaThe culture system contains a layr of amnion membrane as an arterial intima. The methods for collecting, preparing and storing amnion membranes were studied. In addition, the system was further refined to facilitate convenient use. Several types of modified systems were made according to the different purposes.2. The properties of endothelial cells cultured in the 3-D culture systemCompared to endothelial cells cultured in conventiona plastic dish, the endothelial cells cultured in this system bear several properties similar to those endothelial cells in vivo, including the formation of tight junctions, expression of ZO-1 protein, and polarized secretion of endothelin-1.3. Application in atherosclerosis studyIt was found that this system is specifically useful in analyzing monocyte, lymphocyte and endothelial cell interactions, which is one of the earliest process in atherogenesis. We also found that plasm … More a LDL could be modified or oxidized by cultured endothelial cells through which, a fatty streak-like lesion can be produced in vitro when macrophages were co-present. We have attempted to study the influence of various kinds of cytokines such as interleukin-1 and tumor necrosis factor in the fatty streak formation using the current system. To clarify the mechanisms involved in this process, highly purified monocytes, lymphocytes and their subpopulation are required. Currently, we are trying to use immno-magnet isolation method for this purpose.4. Application of the 3-D culture system for non-vascular models-Development of an organ model.A novel type of the 3-D culture system has been developed ; it consists of upper and lower compartments divided by a layr of amnion membrane. On each side of the membrane surface, different types of cells can be cultured separately. In this study, epithelial cells from tracheal mucosa are cultured on the upper surface and endothelial cell or fibroblasts on the lower at the same time. We found that it may be possible to use this 3-D culture system as a new organ model in the future.According the above results, custom-made 3-D culture system of arterial intima or modified 3-D systems for different aims can be easily available commercially from some vendors. We believe that this system is not only useful for the study of atherosclerosis but for the study of inflammation mechanism and cancer metastasis by molecular biology as well. Less