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目的建立胶质疤痕体外模型,观察其形态及细胞外基质的表达情况。方法体外分别培养来自大脑皮层的星形胶质细胞和脑膜成纤维细胞,经胶质纤维酸性蛋白(GFAP)和纤连蛋白(FN)抗体免疫细胞化学染色鉴定后,将两种细胞混合共培养,2d后添加转化生长因子-β1(TGF-β1),以未添加TGF-β1为对照组;GFAP和FN免疫细胞化学染色观察胶质疤痕的形态;免疫细胞化学染色和Western blotting观察促红素肝细胞受体B2(Eph B2)、Eph受体作用配体B2(ephrin B2)、神经蛋白聚糖(neurocan)和神经抗原2(NG2)表达情况。结果疤痕样细胞团簇结构主要是由星形胶质细胞和成纤维细胞共同组成;实验组Eph B2、ephrin B2和neurocan、NG2表达量较对照组明显增加(P<0.05)。结论体外混合培养星形胶质细胞与成纤维细胞并添加TGF-β1后成功建立体外中枢神经系统损伤后疤痕模型。
Objective To establish an in vitro model of glial scar and observe its morphology and extracellular matrix expression. Methods The astrocytes and meningococcal fibroblasts from cerebral cortex were cultured in vitro. After immunofluorescence staining with glial fibrillary acidic protein (GFAP) and fibronectin (FN) antibody, the two cells were co-cultured , TGF-β1 (TGF-β1) was added after 2 days, and TGF-β1 was not added as the control group. GFAP and FN immunocytochemistry staining was used to observe the morphology of glial scar. Immunocytochemical staining and Western blotting were used to observe the expression of erythropoietin Eph B2, ephrin B2, neurocan and NG2. Results The cluster structure of scar-like cells was mainly composed of astrocytes and fibroblasts. The expression of Eph B2, ephrin B2, neurocan and NG2 in the experimental group was significantly higher than that in the control group (P <0.05). Conclusion The in vitro model of ascites and fibroblasts mixed with TGF-β1 was successfully established in vitro.