Griffin S, Clark J B, Canevari L
Department of Molecular Neuroscience, Miriam Marks Division of Neurochemistry, Institute of Neurology, University College London, Queen Square, London, UK.
J Neurochem. 2005 Nov;95(4):1015-22. doi: 10.1111/j.1471-4159.2005.03418.x. Epub 2005 Sep 29.
We looked at the possible interactions between astrocytes and neurones during reperfusion using an in vitro model of ischaemia-reperfusion injury, as a controlled environment that lends itself easily to manipulation of the numerous variables involved in such an insult. We constructed a chamber in which O2 can be lowered to a concentration of 1 microm and developed a primary cortical neuronal culture that is 99% pure and can survive to at least 10 days in vitro. We also established a novel system for the co-culture of astrocytes and neurones in order to study the communication between these cells in a manner that allows the complete separation of one cell type from another. Neurone cultures showed profound cell death following an ischaemic period of only 15 min. We co-cultured neurones that had been subjected to a 15-min ischaemic insult with either non-insulted astrocytes or astrocyte-conditioned medium during the reperfusion stage. Both astrocytes and astrocyte-conditioned medium enhanced neuronal survival. Our data also suggest that astrocyte-sourced neuronal glutathione synthesis may play a role in preventing neuronal death.
我们使用缺血再灌注损伤的体外模型，研究了再灌注期间星形胶质细胞与神经元之间可能的相互作用，该模型提供了一个易于控制的环境，便于对这种损伤中涉及的众多变量进行操作。我们构建了一个能将氧气浓度降至1微摩尔的小室，并培养出了纯度达99%且能在体外存活至少10天的原代皮质神经元。我们还建立了一种星形胶质细胞与神经元共培养的新系统，以便以一种能将一种细胞类型与另一种细胞类型完全分离的方式研究这些细胞之间的通讯。仅缺血15分钟后，神经元培养物就出现了严重的细胞死亡。在再灌注阶段，我们将遭受15分钟缺血损伤的神经元与未受损伤的星形胶质细胞或星形胶质细胞条件培养基进行共培养。星形胶质细胞和星形胶质细胞条件培养基均能提高神经元的存活率。我们的数据还表明，星形胶质细胞来源的神经元谷胱甘肽合成可能在预防神经元死亡中发挥作用。
