The preparation of rat's acellular spinal cord scaffold and co-culture with rat's spinal cord neuron in vitro
The preparation of rat's acellular spinal cord scaffold and co-culture with rat's spinal cord neuron in vitro
复制标题
大鼠脱细胞脊髓支架的制备及与大鼠脊髓神经元的体外共培养
DOI：
10.1038/sc.2016.144
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发表时间：
2017-04-01
期刊：
影响因子：
2.2
通讯作者：
Feng，S-Q
中科院分区：
文献类型：
--
作者：
Ban，D-X;Liu，Y.;Feng，S-Q
实验设计：体外制备大鼠脱细胞脊髓支架（ASCS）与脊髓神经元的生物相容性研究目的：制备ASCS并与神经元共培养，为临床治疗脊髓损伤（SCI）奠定基础。应用苏木精-伊红染色、髓鞘染色和扫描电镜观察ASCS表面结构。结果：制备的ASCS呈网状结构，有大小不一的小孔。H&E染色显示细胞成分全部去除。ASCS具有良好的三维网络多孔结构。DNA琼脂糖凝胶电泳未发现ASCS中的DNA成分。培养的细胞表达神经元特异性烯醇化酶（NSE）抗原与长轴突。H&E染色显示神经元贴附于ASCS孔结构上，细胞生长良好。MTT法检测细胞存活率为（97.53± 1.52）%。免疫组织化学染色显示支架上的神经元表达NSE和NeuN抗原。细胞排列紧密，通道结构充满神经元。结论：ASCS结构完整，神经元在支架内排列紧密。这些结果为临床应用生物材料替代胶质瘢痕治疗脊髓损伤奠定了基础。
Study design:The rat’s acellular spinal cord scaffold (ASCS) and spinal cord neurons were prepared in vitro to explore their biocompatibility.Objectives:The preparation of ASCS and co-culture with neuron may lay a foundation for clinical treatment of spinal cord injury (SCI).Setting:Tianjin Medical University General Hospital, ChinaMethods:ASCS was prepared by chemical extraction method. Hematoxylin and eosin (H&E), myelin staining and scanning electron microscope were used to observe the surface structure of ASCS. Spinal cord neurons of rat were separated in vitro, and then co-cultured with prepared ASCS in virto.Results:The prepared ASCS showed mesh structure with small holes of different sizes. H&E staining showed that cell components were all removed. The ASCS possessed fine three-dimensional network porous structure. DNA components were not found in the ASCS by DNA agarose gel electrophoresis. The cultured cells express neuron-specific enolase (NSE) antigen with long axons. H&E staining showed that the neurons adhered to the pore structures of ASCS, and the cell growth was fine. The survival rate of co-cultured cells was (97.53±1.52%) by MTT detection. Immunohistochemical staining showed that neurons on the scaffold expressed NSE and NeuN antigen. Cells were arranged closely, and the channel structures of ASCS were fully filled with neurons. The cells accumulated in the channel and grew well in good state.Conclusion:The structure of ASCS remained intact, and the neurons were closely arranged in the scaffolds. These results may lay a solid foundation for clinical treatment of SCI when considering glial scar replacement by biomaterials.