两种细胞因子联合诱导骨髓间充质干细胞向神经样细胞分化

Bone marrow mesenchymal stem cells differentiate into neuron-like cells induced by combination of two cytokines

背景:骨髓间充质干细胞可被定向诱导分化为神经样细胞,理论上骨髓间充质干细胞可作为种子细胞应用于周围神经组织工程。目的:用2种细胞因子联合诱导骨髓间充质干细胞分化为神经样细胞,进一步探讨其在周围神经损伤方面的应用。方法:从Wistar大鼠胫骨和股骨提取骨髓间充质干细胞,采用差速贴壁法进行培养和纯化,联合应用碱性成纤维细胞生长因子和表皮生长因子对细胞进行诱导,观察细胞形态变化,并用免疫组织化学方法检测骨髓间充质干细胞中神经元特异性标志物的表达;并研究终止诱导后骨髓间充质干细胞的形态及免疫组织化学方面的变化。结果与结论:骨髓间充质干细胞经诱导后呈典型神经细胞样改变,有两个或多个突起,突起之间相互连接成网状,可见细胞核及核仁,免疫组化检测神经元特异性烯醇化酶、神经丝蛋白及突触素蛋白表达阳性。撤除诱导条件后大部分细胞逐渐恢复原来的成纤维细胞样形态,上述3种蛋白的表达量明显下降。表明应用神经营养因子可诱导骨髓间充质干细胞分化为神经样细胞,但其形态及组织学变化仅能维持一段较短的时间。

BACKGROUND：Bone marrow mesenchymal stem cells （BMSCs） can be induced to differentiate into neuron-like cells directional y. Accordingly, BMSCs can be used as seed cells theoretical y in constructing tissue-engineered peripheral nerves. OBJECTIVE：Using combination of two cytokines to induce BMSCs differentiating into neuron-like cells directional y, and further to discusse its application in peripheral nerve injury. METHODS：BMSCs were isolated and purificated from the bone marrow of Wistar rats by using the differential adherence method. Basic fibroblast growth factor and epidermal growth factor were used to induce the BMSCs differentiating into neuron-like cells. The morphological change was observed and the neuronal specific markers were detected by immunohistochemistry technique. The morphological and immunohistological changes were also studied after the induce agent were removed. RESULTS AND CONCLUSION：With presence of morphological and immunohistochemical features of nerve cells induced by neurotrophic factors, BMSCs exhibited two or more processes that were interconnected as a meshwork;cellnucleus and nucleus could be observed with strong light refraction of cytoplasm. After immunohistochemical staining, neuroln specific enolase, neurofilament protein and synaptophysin protein positive cells were detected. A great amount of cells reversed to their original fibroblast-like morphology, and the expression of the three above-mentioned proteins decreased as the induce agent withdrawn. Our study showed that BMSCs can be induced to differentiate into neuron-like cells, but the transdifferentiation is a short-time reversible phenomenon.