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目的探讨纤维蛋白支架对神经干细胞和星形胶质细胞分化及增殖的影响。方法分别培养胚胎大鼠脊髓来源的神经干细胞和新生鼠脊髓神经胶质细胞,接种于纤维蛋白支架上,同时用多聚赖氨酸修饰的玻片作为对照。于体外培养不同时间后,用神经丝蛋白(NF200)对神经细胞进行免疫荧光染色,测量各复孔(n=4)内NF阳性细胞的突起长度,计算其平均值;用胶质纤维酸性蛋白(GFAP)对胶质细胞进行染色,各复孔(n=4)内统计5个不同视野的胶质细胞总数和GFAP阳性细胞数,计算GFAP阳性细胞相对数量的平均值。比较在纤维蛋白支架和玻片上神经干细胞分化、神经纤维延伸及神经胶质细胞增殖的差异。同时用免疫印迹技术对荧光染色结果进行验证。上述实验各重复3次。结果纤维蛋白支架组的NF阳性纤维明显长于对照组,GFAP阳性星形胶质细胞相对数量明显少于对照组,GFAP的表达水平明显低于对照组。结论纤维蛋白支架可促进神经干细胞向神经细胞分化,并有利于神经纤维的延伸而抑制星形胶质细胞的增殖和成熟。
Objective To investigate the effect of fibrin scaffold on the differentiation and proliferation of neural stem cells and astrocytes. Methods Cultured embryonic rat spinal cord-derived neural stem cells and neonatal rat spinal cord glial cells were inoculated on fibrin scaffolds, and polylysine-modified slides were used as controls. After culturing at different time in vitro, the nerve cells were immunofluoresced with neurofilament protein (NF200) and the length of protuberances of NF-positive cells in each replicate well (n = 4) were measured. The average value of glial fibrillary acidic protein (GFAP) were used to stain the glial cells. The number of glial cells and the number of GFAP positive cells in 5 different fields were counted in each replicate well (n = 4), and the average number of GFAP positive cells was calculated. Differences in neural stem cell differentiation, nerve fiber extension, and glial cell proliferation on fibrin scaffolds and slides were compared. At the same time, immunofluorescence staining was used to verify the results. The above experiments were repeated 3 times each. Results NF - positive fibrin scaffold group was significantly longer than the control group, GFAP positive astrocytes relative number was significantly less than the control group, GFAP expression was significantly lower than the control group. Conclusion Fibrin scaffolds can promote the differentiation of neural stem cells into nerve cells and promote the extension of nerve fibers and inhibit the proliferation and maturation of astrocytes.