中性红比色法检测大鼠小脑神经瘤活细胞  

Neutral red assay for measuring viability of cerebellum neural cells

作　　者：刘锐克[1] 周军兰[1] 梁建辉[1] 

机构地区：[1]北京大学中国药物依赖性研究所,北京100083

出　　处：《中国药理学与毒理学杂志》2003年第1期74-78,共5页Chinese Journal of Pharmacology and Toxicology

摘　　要：目的　探讨中性红比色法测定细胞活性的有效性 ,优化实验参数 ,并初步探讨该方法测定细胞活性的机制。方法　采用中性红比色法、MTT比色法及台盼蓝拒染法测定大鼠小脑神经瘤细胞 (R2细胞 )的活性。结果　①中性红浓度为 10 0mg·L- 1,孵育时间 4h ,溶解液的pH值为 2～ 4 ,细胞密度在每毫升 (0 .5～ 2 4 )× 10 5范围内 ,吸光度值 (A53 0nm)与活细胞数呈正相关 ,是较为合适的实验参数 ;②低温(4℃ )明显抑制R2细胞对中性红的胞吞作用 ,使A53 0nm值降低。结论　中性红比色法灵敏准确 ,结果稳定 ,操作简便 ,可以用于细胞活性和毒性的评价。AIM To examine the validity of neutral red(NR) assay for measuring cell viability and optimize the experimental conditions. METHODS NR assay, MTT assay and cell counting following trypan blue staining were used for the analyses of cell viability. RESULTS ① Optimal experimental parameters in NR assay were: 100 mg·L -1 of NR, 4 h of NR incubation, pH 2-4 of the dissolving solution of NR from R2 cells, and within the range of cells (0.5-24)×10 8·L -1, there was good correlation between NR absorbance (A 530 nm) and viable cell numbers; ② NR endocytosis by R2 cells was very significantly blocked at 4℃. CONCLUSION NR assay is simple, easy and sensitive in determining cell viability and cytotoxicity. The process that NR is taken up by cells may be associated with endocytosis.

关 键 词：细胞毒性 比色法 中性红 胞吞作用 

分 类 号：R739.4[医药卫生—肿瘤]