关键词: 骨髓间充质干细胞, 神经元样细胞, 依达拉奉, 免疫组织化学, 流式细胞术, 扫描电镜, 大鼠

Abstract: Objective To isolate, cultivate and purify rat bone marrow mesenchymal stem cells (MSCs), and induce its directional differentiation into neuron-like cells iEM>n vitro/EM> by basic fibroblast growth factor (bFGF) and edaravone.Methods One month old healthy male Wistar rats were used in this study, MSCs from the bone marrow were isolated and purificated by Percoll medium density gradient centrifugation, repeated subculture and differential adherence methods. After 3 passages, the surface markers of MSCs were detected by immunocytochemistry and flow cytometry. Cell differentiation of MSCs induced by bFGF and edaravone was explored by immunocytochemistry and scanning electron microscopy.Results The cell purity of MSCs was up to 95%. The purified MSCs expressed CD44, but did not express CD34 and CD45. After directional differentiation cultivation, the cells showed the typical neuron appearance cell under the scanning electron microscope and expressed neuron-specific enolase but did not express glial fibrillary acidic protein. Nestin expression in the cells was gradually weakened. BR>Conclusion The results suggest that density gradient centrifugation and differential adhesion methods may be one of the suitable methods of purification of the MSCs, and edaravone can direct and efficiently induce MSCs to differentiate into neuron-like cells. BR>

Key words: Marrow mesenchymai stem cell, Neuron-like cell, Edaravone, Immunohistochemistry, Flow cytometry, Scanning electron microscopy, Rat