中文摘要：

目的 评价脱细胞脊髓支架在大鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells，BMSCs）体外培养中的作用，探索最佳接种浓度。方法 分离、培养及鉴定SD大鼠BMSCs，诱导BMSCs向神经元样细胞分化。制备脱细胞脊髓支架，将第3代BMSCs同脱细胞脊髓支架共培养，MTT法检测细胞增殖活性，比较与普通培养的差异；取第3代BMSCs以不同浓度[（0.5、1、2、3、4）×106/mL]接种于支架（脊髓支架修整0.5 cm小段），检测细胞在支架上的粘附率及上架细胞数，建立接种浓度与细胞粘附率、上架细胞数的关系回归方程；扫描电镜观察脱细胞脊髓支架形态以及细胞与支架的粘附情况。 结果 成功实现BMSCs的分离及培养，BMSCs流式鉴定CD29、CD90阳性表达，向神经元诱导胶质纤维酸性蛋白（GFAP）、巢蛋白（Nestin）呈阳性表达；与普通培养相比，BMSCs在支架上细胞增殖活力显著提高（P〈0.05）；细胞与支架的粘附率在接种浓度为2×106/mL最高，达到（79.6±2.7）%，单位体积上架细胞数达到（1.364±0.047）×106/cm3；扫描电镜观察支架空间结构良好，细胞与支架粘附良好，共培养第3天较第1天细胞数量明显增加。 结论 脱细胞脊髓支架具有多通道的空间结构，适合BMSCs粘附、存活、增殖，该支架是良好的天然脊髓组织工程材料。当粘附率及细胞上架数基本达到最大时的最佳接种浓度为2×106/mL。

英文摘要：

Objective To determine the effect of the acellular scaffold of spinal cord on the rat bone marrow mesenchymal stem cells （BMSCs） cultured in vitro and explore the best inoculation concentration. Methods SD rat BMSCs were separated, cultivated and identified. The obtained BMSCs were induced to differentiate into neurons. The acellular scaffold of spinal cord was prepared. The BMSCs at the third passage were co-cultured with the acellular scaffold of spinal cord. MTT assay was applied to assay the cell proliferation on the BMSCs cultured alone or on the scaffold. The BMSCs at the third passage at different concentrations （0.5, 1, 2, 3, or 4×106/mL） were seeded on the scaffolds （with spinal cord scaffolds trimmed as 0.5 cm small section）. and then the cell adhesion rates and cell numbers on scaffolds were detected to establish a regression equation of inoculation concentration with cell adherence rate and cell numbers on scaffold. Scanning electron microscopy was also used to observe the shape of acellular scaffold of spinal cord and the adhesion between the cells and scaffold. Results BMSCs were successfully isolated and cultivated,with positive expression of CD29 and CD90 detected by flow cytometry. After induction, the cells were positive to glial fibers acid protein （GFAP）and Nestin. Compared with the ordinary plates, BMSCs’ proliferation activity on scaffolds was obviously strengthened （P〈0.05）. Cells adherence rate on scaffolds reached the highest level of （79.6±2.7）% when the cells were seeded at a density of 2×106/mL on the bracket. The maximum cell number per unit volume scaffold was（1.364±0.047）×106/cm3. The adhesion of the cells on the scaffolds was very well under a scanning electron microscope. The number of co-cultivation cells was increased significantly in the third day than in the first. Conclusion The scaffold with multi-channel spatial structure is suitable for BMSCs adhesion, growth, and proliferation. So it is a go