脱细胞神经基质诱导大鼠骨髓间充质细胞向Schwann细胞分化的形态学研究

为探讨脱细胞神经基质移植物(ANA)体外诱导成年大鼠骨髓间充质细胞(BMSCs)向Schwann细胞分化的可行性,本实验将分离的大鼠BMSCs进行体外培养扩增,行表型鉴定后,取第五代细胞,用ANA匀浆进行诱导。诱导后观察细胞的形态变化,免疫细胞化学染色和流式细胞术分别检测细胞中S-100、神经胶质纤维酸性蛋白(GFAP)的表达情况;RT-PCR检测诱导前后细胞的GFAP及S-100的变化;MTT测定不同诱导剂浓度诱导后的细胞活性。结果显示:BMSCs表型鉴定为CD44+、CD54+、CD34-,免疫细胞化学染色GFAP、S-100阳性细胞比例在诱导后分别为(64±5)%、(42±4)%;流式细胞仪检测和RT-PCR反应均显示GFAP、S-100的表达诱导组较对照组有所升高;MTT检测表明诱导剂浓度为1.0mg/ml时诱导后有活性细胞的数目最多。上述结果提示ANA可诱导成年大鼠骨髓间充质细胞分化为Schwann细胞。

MORPHOLOGIC STUDY OF BONE MARROW STROMAL CELLS OF RAT DIFFERENTIATING INTO SCHWANN CELLS BY ACELLULAR NEVER ALLOGRAFTS

To investigate the differentiation of adult rat bone marrow stromal cells(BMSCs) into Schwann cells,BMSCs were induced by acellular nerve allografts(ANA) in vitro.After phenotype characterization,the fifth generation of the rats' BMSCs were extracted,cultured and amplificated in vitro,and then induced by ANA homogenate.The change of cellular morphology was observed and the expression of S-100,glial fibrillary acidic protein(GFAP) were detected in the induced cells with immunocytochemical staining,flow cytometry.The change of GFAP and S-100 in the induced or uninduced cells were detected by RT-PCR.While the cell activity was detected by MTT method.The results showed that the phenotype of BMSCs were CD44+,CD54+and CD34-.About 64%±5% or 42%±4% positive cells in the induced cell were glial fibrillary acidic protein(GFAP) or S-100 immunopositive.By using RT-PCR and flow cytometer,expression of GFAP and S-100 were also detected much higher in induced group than in control group.The MTT detection showed that the number of active cells were maximum when 1.0 mg/ml inductor was applied.The above results indicated that ANA can induce adult rat BMSCs differentiating into Schwann cells.