羧基荧光素琥珀酰亚胺酯标记肿瘤细胞的条件优化  

Optimization of tumor cells labeled with fluorescent dye of CFDA-SE

作　　者：夏爽[1] 秦箐[2,1] 高明星[1] 刘全军[1] 王闻楚[1] 邝晓聪[1] 

机构地区：[1]广西大学化学化工学院,广西南宁530021 [2]广西医科大学药学院天然药物化学教研室,广西南宁530021

出　　处：《现代医药卫生》2011年第5期649-651,共3页Journal of Modern Medicine & Health

基　　金：国家自然科学基金资助项目(项目批准号:30560186);广西科技厅广西科学基金项目(合同编号:桂科自0728128);广西科技厅广西科学基金项目(合同编号:桂科自0991147);广西大型仪器协作共用网资助

摘　　要：目的:探讨化学染料羧基荧光素二乙酸盐琥珀酰亚胺酯(CFDA-SE)标记肿瘤细胞的可行性和最适条件。方法:用不同标记时间、不同浓度、不同培养浓度的CFDA-SE标记肿瘤细胞TCA(舌癌)。结果:20 min内荧光强度与标记时间成正比;荧光强度的持续时间与标记时间成正比,但标记时间超过20 min后,荧光持续时间无明显变化;提高标记时染料浓度,可适当延长荧光持续时间;将试剂盒浓度1/4染料加入培养基中,荧光持续效果最好。结论:CFSE标记肿瘤细胞的影响因素较多,不可用于肿瘤细胞长时间培养的实验;但可用于短时肿瘤细胞标记,确定最佳标记时间为10 min,标记浓度为试剂盒所示浓度,共培养时将试剂盒浓度1/4的染料加入培养基中荧光持续效果最好,且实验操作简单,结果易观察。Objective：To study the feasibility and optimal conditions of chemical dye carboxyfluorescein diacetate succinimidyl ester（CFDA-SE） on labeling tumor cells.Methods：TCA was labeled with CFDA-SE on different labelling time,different concentrations and different culture concentrations.Results：At 20min mark,the fluorescence intensity was proportional to the time;the time duration of fluorescence intensity was proportional to the labeling time.But at the mark for more than 20mim,the fluorescence had no significant change in duration.Increasing the marker dye concentration,the fluorescence duration could be extended.Adding dye with 1/4 concentration of the kit to medium,the fluorescence had best lasting effect.Conclusion：CFDA-SE labelling tumor cells has many influence factors,which can not be used in the tumor cell culture experiments needing long time,but can be used for short-term markers of tumor cells with the optimum marking time for 10min and the labeling concentration shown in the kit.concentration of cultured the concentration of the kit 1/4 medium.In coculture,adding dye with 1/4 of kit concentration to medium could obtain the best fluorescent lasting effect.The experiment is simple to be operated and the results are easy to observe.

关 键 词：CFDA-SE 肿瘤细胞 最适条件 

分 类 号：R392[医药卫生—免疫学]