Investigation for mechanism and protection of retinal ganglion cell death using several culture disease-model systems

体外结果与体外结果不一致并不罕见。培养细胞与体内细胞的环境差异应该是一个可能的原因。然而，与其他类型的细胞一起培养的细胞不适合研究它们对某些负载的纯反应。在本研究中，我们采用纯隔离细胞来识别它们对特定载荷的特定反应，并建立了一些人工重建系统，使用不同类型的纯隔离细胞来清楚地研究它们之间的相互作用。该系统能够在分子水平上分析单一细胞在多种加载条件下的反应和细胞间的反应，并能用于研究纯分离细胞在多种加载条件下的反应。采用DNA芯片技术研究了单纯分离的视网膜神经节细胞（RGC）和视网膜胶质细胞在重力负荷下存活率和mRNA反应的差异。结果表明，两种细胞类型对负荷的反应并不相同。我们还发现，RGC和视网膜神经胶质细胞对缺氧负荷的一氧化氮产生速率不同。我们用单纯分离的RGC和视网膜胶质细胞重建了人工混合培养条件。结果表明，视网膜神经胶质细胞主要对应激负荷产生反应，从而对RGC的存活产生次要影响。这些结果表明，在研究神经保护和神经元死亡机制时，研究细胞间反应是极其重要的。

It is not rare that in vitro results are not consistent in vitro results. The environmental difference of cultured cells from those of in vivo should be one possible reason. However, cells cultured with other types of cells are not suitable for researching their pure reaction against certain loadings. In this study, we employed purely isolated cells for identifying their specific reaction against certain loadings and we established some artificial reconstruction systems using different types of purely isolated cell for investigating their interaction clearly. In conclusion, it is able to analyze single type cell reaction against several loading conditions and intercellular reaction at the molecular level.For studying reactions of purely isolated cells against several loading conditions we developed a novel quantitative gravity loading system. We investigated differences between purely isolated retinal ganglion cell (RGC) and retinal glial cell against gravity loading in their survival rate and in mRNA reaction employing DNA chip method. In results, the reactions to loading were not the same between two cell types. We also revealed that nitric oxide producing rate against hyoxic loading are different between RGC and retinal glial cells. We reconstructed artificial mixed culture condition using purely isolated RGC and retinal glial cells. In results, retinal glial cells primary reacted stress loading resulting secondary effect on RGC survival. These results indicate that it is extremely important to investigate intercellular reaction when investigating neuroprotection and neuronal death mechanism.