目的探讨应用量子点(Quantum dots,Qdots)体外标记胚胎干细胞进行长期跟踪的可行性。方法以Qtracker 655标记小鼠未分化胚胎干(ES)细胞和小鼠胚胎成纤维(MEF)细胞,标记后倒置荧光显微镜和流式细胞仪动态观察细胞内Qdots的变化。丝裂霉素、秋水仙素或Verapamil处理标记Qdots的胚胎干细胞,观察细胞内Qdots含量与细胞增殖及多药耐药泵的关系。结果Qdots可以标记ES细胞,标记效率可达到90%以上。标记24h后细胞内Qdots含量较MEF细胞明显下降,至72h仅5%左右的ES细胞Qdots标记呈阳性。细胞增殖抑制实验显示,ES细胞内Qdots含量下降与细胞增殖关系不大。用Verapamil抑制多药耐药泵的功能,也不能阻止胞内Qdots含量的降低。结论Qdots不能长期标记未分化ES细胞,ES细胞内Qdots含量的迅速降低并非单由细胞增殖过快造成,而且与迅速地被排出细胞有关,但这一排出与多药耐药泵无关。
Objective The aim of this study is to investigate the feasibility of labeling embryonic stem cells with quantum dots (Qdots) for long-term tracking in vitro. Methods Mouse embryonic stem cells (ES) and mouse embryonic fibroblasts (MEF) were labeled with Qtracker 655. After labeling, intra-cellular Qdots was detected by fluorescence microscope and flow cytometry. The inhibition of cell proliferation was carried out by mitomycin C or colchicines treatment after cell labeling, and the inhibiting of multi-drug ...