[目的]研究慢病毒载体(lentiviral vectors,LVs)介导增强型绿色荧光蛋白(enhanced green fluorescent protein,eGFP)标记大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)的可行性及对细胞生物学特性的影响。[方法]采用密度梯度离心法分离培养大鼠BMSCs,经Lenti-emGFP/hygro慢病毒载体转染和潮霉素药筛,镜下观察细胞形态和eGFP表达情况,MTT法检查转染前后细胞增殖活力,流式细胞仪检测细胞表面标记物(CD34、CD44、CD45、CD90)和eGFP,检测转染后细胞的成骨、成脂分化潜能并分别行茜素红染、油红O染色鉴定。[结果]大鼠BMSCs呈成纤维细胞样形态,漩涡样状排列,转染后细胞形态不变,并高效表达eGFP,两组细胞增殖活力一致,流式细胞仪检测显示,转染后的P3代大鼠BMSCs-eGFP的eGFP阳性率为99.97%,转染前后,两组细胞CD34和CD45阳性率低于2%,而CD44和CD90阳性率高于99%。转染后的P3代大鼠BMSCs-eGFP成骨诱导后,茜素红染色,可见呈桔红色的块状钙结节,成脂诱导后,油红O染色显示有大量红色脂质沉淀,成骨成脂诱导过程中,eGFP持续表达。[结论]Lenti-emGFP/hygro转染大鼠BMSCs并经潮霉素药筛后,eGFP可稳定高效表达,对细胞的生物学特性无明显影响,是一种较为理想的标记方法。 [Objective] To investigate the feasibility of lentiviral vectors (LVs) -mediated enhanced green fluorescent protein (eGFP) labeling of rat bone marrow mesenchymal stem cells (BMSCs) Effects of cell biology. [Methods] Rat BMSCs were isolated and cultured by density gradient centrifugation. Lenti-emGFP / hygro lentiviral vector and hygromycin drug were used to screen the morphology of cells and the expression of eGFP. MTT was used to examine the proliferation of BMSCs before and after transfection (CD34, CD44, CD45, CD90) and eGFP were detected by flow cytometry. The osteogenic and adipogenic potential of transfected cells were detected and stained with Alizarin Red and Oil Red O respectively. [Results] Rat BMSCs were fibroblast-like and arranged in a swirling pattern. After transfection, the morphology of BMSCs was unchanged and eGFP was highly expressed. The proliferation activity of BMSCs in both groups was consistent. Flow cytometry showed that P3 The positive rate of eGFP in rat BMSCs-eGFP was 99.97%. Before and after transfection, the positive rate of CD34 and CD45 in both groups was less than 2%, but the positive rates of CD44 and CD90 were higher than 99%. After induction of osteogenic induction of BMSCs-eGFP in P3 generation, alizarin red staining showed massive orange nodules. After adipogenic induction, oil red O staining showed a large amount of red lipid deposition During adipogenic induction, eGFP is continuously expressed. [Conclusion] The eGFP can be stably and efficiently expressed in BMSCs transfected with Lenti-emGFP / hygro and hygromycin, and has no significant effect on the biological characteristics of cells. It is an ideal labeling method.