转化生长因子β和神经生长因子联合诱导成年鼠骨髓基质细胞分化为神经细胞的研究

Differentiation of bone marrow stroma cells in adult rats into nerve cells induced by transforming growth factor beta and nerve growth factor

目的:探讨转化生长因子β(TGF-β)和神经生长因子联合诱导成年大鼠骨髓基质细胞成神经元样细胞的可能性,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2h先用含1μg/L转化生长因子及200mL/L胎牛血清的DMEM培养液培养,以促进细胞分裂,然后用神经生长因子(NGF)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神经元中间丝蛋白(NF),神经元特异性烯醇化酶(NSE),神经胶质纤维酸性蛋白(GFAP)等特异性标志物。结果:诱导后2h,部分细胞的形态已发生改变,12h后大部分细胞不仅在形态上表现为神经元样特征,而目NF,NSE等特异性抗体呈阳性表达,而GFAP抗体表达呈阴性。结论:TGF-β和NGF能将骨髓基质细胞诱导成神经元样细胞。

AIM:To explore the possibility of transforming growth factor beta(TGF β)and nerve growth factor(NGF) to induce bone marrow stroma cells(BMSCs) into neurons in adult rats so as to provide a new method for neural regeneration and the treatment of spine cord injury. METHODS:BMSCs from adult rats were isolated and cultured for 5 passages.BMSCs were cultured in DMEM containing 200 mL/L fetal bovine serum and 1 μg/L TGF β for 2 hours in order to promote cell division.NGF was used as inducers.The morphological changes of the cells were observed. The expressions of neuron intermediate filament(NF), neuron specific enolase(NSE),glial fibrillary acidic protein(GFAP)and other specific antibodies were detected with immunohistochemical method. RESULTS:After 2 hours of induction, some cells showed morphological changes;12 hours later,most BMSCs became neuron like cells and were expressed positively with NF and NSE specific antibody,but negatively with GFAP. CONCLUSION:BMSCs can be induced into neuron like cells by TGF β and NGF.