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

1.模拟动脉内膜的三维培养系统的建立该培养系统含有一层羊膜作为动脉内膜。对羊膜的收集、制备和保存方法进行了研究。此外，该系统还进一步完善，以方便使用。根据不同的目的制作了几种不同类型的改装系统。与常规塑料培养的内皮细胞相比，三维培养系统培养的内皮细胞具有与体内内皮细胞相似的一些特性，包括紧密连接的形成、ZO-1蛋白的表达和ET-1.3的极化分泌。在动脉粥样硬化研究中的应用研究发现，该系统特别适用于分析单核细胞、淋巴细胞和内皮细胞的相互作用，这是动脉粥样硬化形成最早的过程之一。我们还发现血浆中的…培养的内皮细胞可以修饰或氧化更多的低密度脂蛋白，在巨噬细胞共存的情况下，通过这种修饰或氧化可以在体外产生脂肪条状病变。我们试图利用目前的系统研究各种细胞因子，如白介素1和肿瘤坏死因子在脂肪条纹形成中的影响。为了阐明这一过程中涉及的机制，需要高纯度的单核细胞、淋巴细胞及其亚群。目前，我们正在尝试使用免疫磁体隔离方法来达到这一目的。三维培养系统在非血管模型中的应用-器官模型的发展。一种新型的三维培养系统已经被开发出来；它由被一层羊膜隔开的上下两个隔室组成。在膜表面的每一侧，可以分别培养不同类型的细胞。在本研究中，气管黏膜上皮细胞在上表面培养，内皮细胞或成纤维细胞在下表面同时培养。根据上述结果，定制的动脉内膜三维培养系统或改良的不同用途的三维培养系统可以很容易地从一些厂商那里获得商业上的应用。我们认为，该系统不仅适用于动脉粥样硬化的研究，也有助于从分子生物学的角度研究炎症机制和肿瘤转移。较少

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