TGF-β与BDNF联合诱导成年大鼠BMSC向神经样细胞分化的研究.docVIP

TGF-β和BDNF联合诱导成年大鼠BMSC向神经样细胞分化的研究 （作者:___________单位: ___________邮编: ___________） 作者：梅晰凡，吕刚，王岩松，李全双，郭占鹏 【摘要】 目的 探讨转化因子β(TGF-β)和脑源性神经生长因子(BDNF)联合诱导成年大鼠骨髓基质细胞成神经细胞的可能性，为神经再生及脊髓损伤的治疗提供一种新方法。方法 从大鼠骨髓中分离培养骨髓基质细胞并传至第5代，诱导前2 h先用含1 μg/L转化因子（TGF-β）及20%FBS的DMEM培养液培养，以促进细胞分裂，然后用脑源性神经生长因子(BDNF)作诱导剂，观察细胞形态的变化，并采用免疫组织化学法检测诱导后的细胞表达NF，NSE，GFAP等特异性标志物。结果 诱导后5 h，部分细胞的形态已发生改变，12 h后大部分细胞不仅在形态上表现为神经元样特征，而目NF及NSE等特异性抗体呈阳性表达，而GFAP抗体表达呈阴性。结论 转化因子β(TGF-β)和脑源性神经生长因子(BDNF)能将骨髓基质细胞诱导成神经元样细胞。 【关键词】 骨髓基质细胞；转化因子β；脑源性神经生长因子；细胞分化；神经元样细胞 Abstract：Objective To study the possibility that transforming growth factor-β and brain-derived neurotrophic factor induces adult rat bone marrow stromal cells(BMSCs) into neurons, and provide a new method for neuron regeneration and the treatment of spine cord injury. Methods The bone marrow stromal cells from adult rats were isolated and cultured for 5 passages. The BMSCs were incubated with DMEM containing 20% FRS and TGF-β(1μg/L )for 2 hours. Then the media were replaced by DMEM media consisting of BDNF(50μg/mL). The morphological changes of the cells were observed under phase contrast microscope，the cells were stained immunocytochemically with NF，NSE and GFAP antibodies respectively. Results After 5 hour of induction with neurotrophic factor, some cells showed morphological changes, and 12 hours later,many BMSCs became neuron-like cells and were stained positively with NF，NSE antibody，but negatively with GFAP.Conclusions The bone marrow stromal cells can differentiate into neuronlike cells by the induction of transforming growth factor-βand brain-derived neurotrophic factor. Key words：bone marrow stromal cells；transforming growth factor-β；brain-derived neurotrophic factor；cell differentiation induction；neuron-like cells 近年来的基础研究发现脊髓损伤后采用组织工程方法，将有活性的种子细胞移植治疗脊髓损伤，能促使损伤轴突修复、再生和恢复脊髓部分神经功能，骨髓基质细胞（MSC）可为基因治疗神经系统的疾病提供良好的载体［1］。骨髓基质细胞诱导分化为成骨细胞、脂肪细胞及神经细胞的研究也见报道，为了探讨转化因子β和脑源性神经营养因子对骨髓基质细胞向神经细胞联合诱导分化的可能性和条件，为以后神经细胞再生和脊髓内移植的研究奠定基础［2－3］，本研究观察了转化因子β（transforming