Yu Qin, Luo Yi, E Yan, Sheng Li-xian, Dong Qin, Dong Wei, Guo Ying
Zhejiang TCM College, Hangzhou 310053.
Zhongguo Zhong Xi Yi Jie He Za Zhi. 2005 Jan;25(1):49-53.
To study the effect of Danshensu (DSS) on directional differentiation of mesenchymal stem cells (MSC) into neuron-like cells.
MSC were separated from bone marrow with density gradient centrifugation, wall sticking screening and amplified in vitro. Flow cytometry was used to monitor the expression of surface antigens. DSS contained in non-serum L-DMEM was used to induce differentiation of MSC to neuronlike cells, and the effect of DSS when different concentration and acting time used was explored. And levels of neuron-specific enolase (NSE), neurofilament protein (NF-M), nestin, and expression of glial fibrillary acidic protein (GFAP) were measured by immunohistochemical method.
After being propagated and amplified in vitro, MSC were positively expressed for CD29, CD44, CD166, and negatively expressed for CD14, CD34, CD45, HLA-DR. After induction of DSS, MSC exhibited the typical form of perikaryon with pyknotic cell body and prominence projected like that of neuron. These cells were positively expressed in NSE, NF-M and nestin, and negatively expressed in GFAP.
DSS could induce differentiation of MSC to neuron-like cells in vitro, the action is concentration- and time-dependent.
研究丹参素（DSS）对间充质干细胞（MSC）向神经元样细胞定向分化的影响。
采用密度梯度离心法从骨髓中分离MSC，通过贴壁筛选法进行体外扩增。运用流式细胞术监测表面抗原的表达。采用不含血清的L-DMEM培养基中含有的DSS诱导MSC向神经元样细胞分化，探讨不同浓度及作用时间的DSS的作用效果。采用免疫组织化学方法检测神经元特异性烯醇化酶（NSE）、神经丝蛋白（NF-M）、巢蛋白的水平及胶质纤维酸性蛋白（GFAP）的表达。
体外传代扩增后的MSC，CD29、CD44、CD166呈阳性表达，CD14、CD34、CD45、HLA-DR呈阴性表达。经DSS诱导后，MSC呈现出典型的神经元胞体形态，胞体固缩，有突起伸出。这些细胞NSE、NF-M和巢蛋白呈阳性表达，GFAP呈阴性表达。
DSS能在体外诱导MSC向神经元样细胞分化，其作用具有浓度和时间依赖性。
