在动物模型脏器中动态追踪荧光染料标记的目标细胞  

作　　者：袁璐萍 马炬明[2] 

机构地区：[1]杭州萧山区第一人民医院,310000 [2]杭州中国人民解放军第117医院,310013

出　　处：《浙江临床医学》2015年第10期1703-1705,共3页Zhejiang Clinical Medical Journal

基　　金：浙江省年度重大科技专项计划项目（2011C130291-1）

摘　　要：目的 利用染料羧基荧光素二乙酸盐琥珀酰亚胺酯（CFDA-SE）标记细胞,通过测定荧光信号强度计算细胞数量,作为追踪移植细胞进入模型动物体内后分布情况的方法.方法 采用CFDA-SE标记不同类细胞,荧光显微镜计数染色率,台盼蓝计数存活率,酶标仪检测荧光信号,确定不同种类、同种细胞不同状态下及模拟动物体内细胞数与荧光信号之间的线性关系,制作标准曲线,确定利用该染料作细胞计数的可行性.将CFDA-SE标记细胞经脾脏注入小鼠体内,于3h、6h、12h、24h处死小鼠,检测肝脏信号,根据标准曲线定量计算肝脏内该细胞动力学特征.结果 CFDA SE括记细胞后,荧光信号与细胞数相关性较好,24h内随时间增加荧光信号有所增高,但与细胞数相关性不受影响.加入小鼠内脏组织匀浆后,荧光信号值有所降低,但与细胞数的相关性不受影响.结论 CFDA-SE可标记活细胞,操作简单,标记率高,注入体内后,通过严格制作标准曲线,检测脏器荧光信号,可定量确定细胞在体内脏器的的动态分布,方法简单可靠.Objective To develop a simple method for dynamic tracking and quantitative analysis of transplanted cells inside the body of animal experimental. Method Cells were labeled with Carboxyfluorescein diacetate, succinimidyl ester （CFDA-SE） and the fluorescent signal were measured using. The relationship between cell number and the signal intensity of different cell lines were investigated, and the cell amount within an organ can thus be calculated. The method was verified in animal model. CFDA-SE labeled A549 and HepG2 cells were injected into the spleen of mouse model of acute liver injury induced with CC14. The fluorescent signal were measured at 3, 6, 12, 24 hours after cell injection. The feature of transplanted cell distribution was determined by calculating the percentage of CFDA-SE stained cell retained within the liver. Result There was very good linear correlation between signal intensity and cell number. For a single cell, the fluorescent signal would keep on increasing during the first 24 hours, but the correlation between signal and cell numbers would not be affected. After the stained cells were mixed with tissue homogenate, the signal could be inhibited in some degree, but the correlation between signal intensity and cell numbers would still not be affected. Conclusion The cell number can be calculated using the fluorescent signal of the CFDA-SE labeled cells within a special organ in animal model. It is a simple method with high accuracy for monitoring the dynamic distribution of transplanted cell in the organs of animal model.

关 键 词：细胞示踪 羧基荧光素二乙酸盐琥珀酰亚胺酯 荧光信号 

分 类 号：R730.5[医药卫生—肿瘤]