CAJ | 학술논문

目的：对大鼠骨髓间充质干细胞（bone marrow mesenchymal stem cells ，BMSCs）进行分离、培养与鉴定，并探讨全反式维甲酸（retinoic acid，RA）、碱性成纤维细胞生长因子（fibroblast growth factor，basic， bFGF）和表皮生长因子（ epidermal growth factor ， EGF ）联合诱导BMSCs分化为神经细胞的可行性。方法全骨髓贴壁法分离培养BMSCs ，观察细胞形态及生长增殖情况；流式鉴定细胞表面标志物CD29、CD34、CD90；选用第3代细胞，经RA、bF-GF和EGF联合诱导后，细胞免疫化学染色检测神经细胞标志物神经元特异性烯醇化酶（ neuron specific enolasen ， NSE）的表达。结果体外培养的BMSCs呈成纤维细胞样，第3、4、5代BMSCs的生长曲线均呈S形，活性无明显差异。 BMSCs的均一性较好，第3代细胞CD29、CD90阳性率均在90％以上，而CD34阳性率仅为0．58％；BMSCs经诱导后分化为神经细胞，并表达神经细胞标志NSE。结论成功建立BMSCs 的体外培养体系，所得细胞纯度高、生物学特征稳定，并可诱导分化为神经细胞，为移植治疗神经系统损伤提供实验基础。
목적：대대서골수간충질간세포（bone marrow mesenchymal stem cells ，BMSCs）진행분리、배양여감정，병탐토전반식유갑산（retinoic acid，RA）、감성성섬유세포생장인자（fibroblast growth factor，basic， bFGF）화표피생장인자（ epidermal growth factor ， EGF ）연합유도BMSCs분화위신경세포적가행성。방법전골수첩벽법분리배양BMSCs ，관찰세포형태급생장증식정황；류식감정세포표면표지물CD29、CD34、CD90；선용제3대세포，경RA、bF-GF화EGF연합유도후，세포면역화학염색검측신경세포표지물신경원특이성희순화매（ neuron specific enolasen ， NSE）적표체。결과체외배양적BMSCs정성섬유세포양，제3、4、5대BMSCs적생장곡선균정S형，활성무명현차이。 BMSCs적균일성교호，제3대세포CD29、CD90양성솔균재90％이상，이CD34양성솔부위0．58％；BMSCs경유도후분화위신경세포，병표체신경세포표지NSE。결론성공건립BMSCs 적체외배양체계，소득세포순도고、생물학특정은정，병가유도분화위신경세포，위이식치료신경계통손상제공실험기출。
Objective To isolate and culture rat bone marrow mesenchymal stem cells (BMSCs) and induce them differen-tiate to neural cells .Methods BMSCs were isolated from rats by wall sticking method .Then the cells were identified with morphology , proliferation and the surface markers .Neuron-like cells from BMSCs were induced by the combination of ret-inoic acid(RA),fibroblast growth factor, basic (bFGF)and epidermal growth factor (EGF), and their neuron specific eno-lasen ( NSE ) were detected by NSE immunohistochemistry .Results Cultured BMSCs like fibroblast -shaped in vitro. Growth curves of the 3rd, 4th and 5th-generation BMSCs and their activities were not significantly different .FCM detected that CD29 and CD90 were positively expressed , but CD34 was negatively expressed .The induced BMSCs showed neuron -like morphology and expressed NSE , the neuron marker .Conclusion We have established an efficient and steady method to obtain BMSCs .BMSCs could differentiated into neurons which was an ideal source for clinical cell transplantation in treat -ment of nervous system diseases .