Department of Neurology, Medical Faculty, Heinrich Heine University, 40225 Düsseldorf, Germany.
Molecular Physiology, Center for Integrative Physiology and Molecular Medicine, University of Saarland, 66424 Homburg, Germany.
Int J Mol Sci. 2023 Jun 30;24(13):10972. doi: 10.3390/ijms241310972.
Stroke is a major reason for persistent disability due to insufficient treatment strategies beyond reperfusion, leading to oligodendrocyte death and axon demyelination, persistent inflammation and astrogliosis in peri-infarct areas. After injury, oligodendroglial precursor cells (OPCs) have been shown to compensate for myelin loss and prevent axonal loss through the replacement of lost oligodendrocytes, an inefficient process leaving axons chronically demyelinated. Phenotypic screening approaches in demyelinating paradigms revealed substances that promote myelin repair. We established an ex vivo adult organotypic coronal slice culture (OCSC) system to study repair after stroke in a resource-efficient way. Post-photothrombotic OCSCs can be manipulated for 8 d by exposure to pharmacologically active substances testing remyelination activity. OCSCs were isolated from a NG2-CreERT2-td-Tomato knock-in transgenic mouse line to analyze oligodendroglial fate/differentiation and kinetics. Parbendazole boosted differentiation of NG2 cells and stabilized oligodendroglial fate reflected by altered expression of associated markers PDGFR-α, CC1, BCAS1 and Sox10 and GFAP. In vitro scratch assay and chemical ischemia confirmed the observed effects upon parbendazole treatment. Adult OCSCs represent a fast, reproducible, and quantifiable model to study OPC differentiation competence after stroke. Pharmacological stimulation by means of parbendazole promoted OPC differentiation.
中风是由于再灌注以外的治疗策略不足导致持续残疾的主要原因，导致少突胶质细胞死亡和轴突脱髓鞘，梗死周围区域持续炎症和星形胶质细胞增生。损伤后，已证明少突胶质前体细胞 (OPC) 通过替代丢失的少突胶质细胞来补偿髓鞘丢失并防止轴突丢失，这是一个效率低下的过程，导致轴突长期脱髓鞘。脱髓鞘范式中的表型筛选方法揭示了促进髓鞘修复的物质。我们建立了一种体外成年冠状切片培养 (OCSC) 系统，以经济高效的方式研究中风后的修复。通过暴露于测试髓鞘再生活性的药理活性物质，可以对光血栓后 OCSC 进行 8 天的操作。OCSC 是从 NG2-CreERT2-td-Tomato 基因敲入转基因小鼠系中分离出来的，用于分析少突胶质细胞命运/分化和动力学。帕苯达唑促进了 NG2 细胞的分化，并通过改变相关标记物 PDGFR-α、CC1、BCAS1 和 Sox10 和 GFAP 的表达稳定了少突胶质细胞命运。体外划痕实验和化学性缺血证实了帕苯达唑处理后的观察到的效果。成年 OCSC 代表了一种快速、可重复和可量化的模型，可用于研究中风后 OPC 分化能力。通过帕苯达唑的药理刺激促进了 OPC 的分化。
