CAJ | 학술논문

背景:骨髓间充质干细胞定向分化为神经干细胞并探讨提高分化效率的方法,具有重要的理论与实际应用意义.目的:建立以不同诱导方法横向分化大鼠骨髓间充质干细胞为功能神经元的方法.方法:分离、纯化得到骨髓间充质干细胞,流式细胞仪检测骨髓间充质干细胞表面标志物.将骨髓间充质干细胞分为神经营养因子诱导组、化学诱导组和对照组,骨髓间充质干细胞分别加入脑源性神经营养因子和碱性成纤维细胞生长因子、二甲亚砜和丁香茴醚、PBS进行诱导.结果与结论:神经营养因子诱导组和化学诱导组诱导后的细胞均表达神经元特异性烯醇化酶和微管相关蛋白2,且2组细胞的表达量接近(P>0.05),而对照组未发现神经元特异性烯醇化酶和微管相关蛋白2阳性表达.膜片钳系统检测发现神经营养因子诱导组诱导后的细胞具有神经细胞特征性的动作电位和兴奋性突触后电流,而化学诱导组和对照组均未发现动作电位和兴奋性突触后电流.提示脑源性神经营养因子联合碱性成纤维细胞生长因子是一种诱导骨髓间充质干细胞横向分化为功能神经元的可靠方法.
배경:골수간충질간세포정향분화위신경간세포병탐토제고분화효솔적방법,구유중요적이론여실제응용의의.목적:건립이불동유도방법횡향분화대서골수간충질간세포위공능신경원적방법.방법:분리、순화득도골수간충질간세포,류식세포의검측골수간충질간세포표면표지물.장골수간충질간세포분위신경영양인자유도조、화학유도조화대조조,골수간충질간세포분별가입뇌원성신경영양인자화감성성섬유세포생장인자、이갑아풍화정향회미、PBS진행유도.결과여결론:신경영양인자유도조화화학유도조유도후적세포균표체신경원특이성희순화매화미관상관단백2,차2조세포적표체량접근(P>0.05),이대조조미발현신경원특이성희순화매화미관상관단백2양성표체.막편겸계통검측발현신경영양인자유도조유도후적세포구유신경세포특정성적동작전위화흥강성돌촉후전류,이화학유도조화대조조균미발현동작전위화흥강성돌촉후전류.제시뇌원성신경영양인자연합감성성섬유세포생장인자시일충유도골수간충질간세포횡향분화위공능신경원적가고방법.
BACKGROUND:It has the important theoretical and practical significance to induce the differentiation of bone marrow mesenchymal stem cel s into neural stem cel s and to investigate the method to improve the differentiation efficiency. OBJECTIVE:To investigate the method of trans-differentiation of bone mesenchymal stem cel s into functional neurons. METHODS:Bone marrow mesenchymal stem cel s were isolated and purified from rat bone marrow by density gradient centrifugation and adherent culture. Expression of bone marrow mesenchymal stem cel s surface marker was detected by flow cytometry. Al bone marrow mesenchymal stem cel s were divided into three groups:neurotrophic factor induction group, chemical induction group and control group. The bone marrow mesenchymal stem cel s in the neurotrophic factor induction group were induced with brain-derived neurotrophic factor and basic fibroblast growth factor, the cel s in the chemical induction group were induced with dimethyl sulfoxide and butylated hydrochloride, and the cel s in the control group were induced with PBS. RESULTS AND CONCLUSION:The bone marrow mesenchymal stem cel s in the induction groups could express neuron specific enolase and microtubule-associated protein-2, and there was no significant difference of the expression rate between these two groups (P>0.05), while in the control group, no expression of neuron specific enolase and microtubule-associated protein-2 was detected. Patch clamp system detection showed that the induced bone marrow mesenchymal stem cel s in the neurotrophic factor induction group could express action potential with the characteristics of neural cel s and excitatory postsynaptic currents. There were no electrophysiologica characteristics in the chemical induction group and control group. It indicates that brain-derived neurotrophic factor combined with basic fibroblast growth factor is an effective method of inducing bone marrow mesenchymal stem cel s to trans-differentiate into functional neurons.