摘要
目的观测转染增强型绿色荧光蛋白基因(enhanced green fluorescent protein gene,EGFP)并稳定表达前后兔骨髓间充质干细胞(bone marrowmesenchymal stem cells,BMSCs)的生物学特性,探讨利用EGFP作为体内试验示踪剂的可行性。方法利用pLEGFP-N1逆转录病毒载体质粒转染PT67包装细胞,提取病毒上清液后感染BMSCs,G418筛选及单克隆培养得到稳定表达绿色荧光蛋白的BMSCs细胞。比较转染前后BMSCs的生长曲线,细胞活性和贴壁率。结果获得了稳定表达绿色荧光蛋白的BMSCs,其生物学特性无明显改变,荧光有效表达时间达3个月。结论利用逆转录病毒法转染EGFP可作为BM-SCs体内示踪。
Objective Stably introduce enhanced green fluorescent protein gene （EGFP） to bone marrow mesenchymal stem cells （BMSCs） and investigate the change of biological characteristics. To approach the feasibility of EGFP he applied as a stable morphological marker for BMSCs in vivo. Methods Packaging cells PT67 were transfected by pLEGFP-N1 retroviral vector,and BMSCs were transduced by viral supernatant infection. BMSCs stably express EGFP （EGFP-BMSCs） was obtained by be selected with G418 and single cell cloning. EGFP-BMSCs was compared with BMSCs in growth curve , cytoaetive and adherence rate. Results EGFP was stably introduced to BMSCs by retroviral vector. The biological characteristics of EGFP-BMSCs were consistent with those of BMSCs and fluorescent light expression can persist three months. Conclusions EGFP can be applied as a stable morphological marker for BMSCs in VlVO.
出处
《口腔生物医学》
2010年第1期29-33,共5页
Oral Biomedicine
