氯甲基苯甲酰氨和5-乙炔基-2’-脱氧尿嘧啶核苷标记示踪大鼠脂肪干细胞的研究  

Chloromethyl - and 5-ethynyl-2＇ -deoxyuridine tracker labeling for rat adipose-derived stem cells

作　　者：转黎 王涛[1] 李明超[1] 张岩[1] 刘继红[1] 叶章群[1] 

机构地区：[1]华中科技大学同济医学院附属同济医院泌尿外科,武汉430030

出　　处：《中华实验外科杂志》2013年第8期1701-1703,共3页Chinese Journal of Experimental Surgery

基　　金：国家自然科学基金资助项目（81070484）

摘　　要：目的比较研究氯甲基苯甲酰氨（CM-Dil）和5-乙炔基-2＇-脱氧尿嘧啶核苷（EdU）对大鼠脂肪干细胞（ADSCs）的体外标记示踪效果，为ADSCs移植治疗勃起功能障碍体内研究寻找最佳的干细胞示踪剂。方法分离和培养sD大鼠ADSCs，分别采用10μmol／LCM-Dil荧光活性染料和50μmol／LEdU标记第4代ADSCs，并通过流式细胞术和荧光显微镜检测标记后第7、14、21、28天细胞阳性率。结果CM-Dil和EdU两者对ADSCs生物学特性均无明显影响。CM-Dil标记ADSCs阳性率达（99．6±0．2）％，第21天后荧光开始衰减，第35天细胞阳性率仍达（64．2＋3．3）％；EdU标记ADSCs阳性率为（87．1±1．8）％，7d后细胞阳性率逐渐降低，第14天后急剧降低，第28天细胞阳性率为（14．4±2．7）％。结论CM-Dil和EdU两者均适用于ADSCs标记示踪，CM-Dil标记具有操作简单、标记阳性率高、荧光持久等优点，适用于长期标记示踪，EdU仅适用于短期标记示踪。Objective To compare the labeling effects of chloromethyl-benzamidodialkylcarboeya- nine （CM-Dil） vs. 5-ethynyl-2＇ -deoxyuridine （EdU） on rat adipose-derived stem cells （ ADSCs ）, and seek the optimal stem cell labeling technique applied to erectile dysfunction （ED） therapy. Methods Rat ADSCs were isolated and cultured from the adipose tissue of inguinal region. The ADSCs at the fourth pas- sage were labeled with 10 μmol/L CM-Dil or 50 μ mol/L EdU, and the positive cell rate and fluorescence intensity were detected by using flow cytometry and fluorescence microscope at 7th, 14th, 21st and 28th day, respectively. Results CM-Dil and EdU had no effects on the proliferation of ADSCs. The positive rate of CM-Dil labeling was （99. 6±0. 2）%, and fluorescence decay began at 21st day, and the positive rate was （ 64. 2 ± 3.3 ） % at 35th day. The positive rate of EdU labeling was （ 87. 1 ± 1.8 ） %, and de- creased at 7th day, and the positive rate was only （ 14. 4 ± 2. 7） % at 28th day. Conclusion CM-Dil and EdU were suitable for ADSCs tracker labeling. However, CM-Dil gains much more advantages than EdU in long-term tracing, which may be an ideal ADSCs tracing technology in the future treatment of ED.

关 键 词：氯甲基苯甲酰氨 5-乙炔基-2’-脱氧尿嘧啶核苷 示踪 脂肪干细胞 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]