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的情况并不罕见。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。在这项研究中，我们使用纯分离细胞来鉴定它们对某些负载的特异性反应，并使用不同类型的纯分离细胞建立了一些人工重建系统来清楚地研究它们的相互作用。总之，它能够在分子水平上分析多种负载条件下的单型细胞反应和细胞间反应。为了研究纯分离细胞在几种负载条件下的反应，我们开发了一种新的定量重力加载系统。采用DNA芯片法研究了纯分离的视网膜神经节细胞（RGC）和视网膜胶质细胞在重力载荷下的存活率和mRNA反应的差异。结果表明，两种细胞类型对负载的反应是不一样的。我们还发现，RGC细胞和视网膜胶质细胞对缺氧负荷产生一氧化氮的速率不同。我们用分离的RGC细胞和视网膜胶质细胞重建人工混合培养条件。结果表明，视网膜胶质细胞主要反应应激负荷，从而对RGC存活产生继发性影响。这些结果表明，在研究神经保护和神经元死亡机制时，研究细胞间反应是非常重要的。

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.