激光生物学报2011，Vol.20Issue(2)：269-273,5.DOI:10.3969/j.issn.1007-7146.2011.02.023
贴壁培养细胞台盼蓝拒染试验的方法学探讨
A Methodological Study of Trypan Blue Exclusion Test for Cultured Adherent Cells
摘要
Abstract
The present work aimed at making an approach to the trypan blue exclusion test (TBET) for cultured adherent cells, and offering suggestions for its adequate application.Human retinal pigment epithelial cell line-19 (ARPEl9) was used in this work.The cells were cultured in growth medium with or without the addition of sodium azide ( NaN3 ).The survival percentage of attached cells was measured by in situ TBET as well as hemacytometer-based one.The latter one was performed for detached cells in the culture supernatant.Similar to the knowledge of hemaeytometerbased TBET, measured values of percent cell survival by in situ TBET increased gradually with the increase in either dye concentration or staining time.The survival percentages of detached cells were markedly lower than those of attached cells.The number of detached cells increased extensively owing to NaN3 exposure.Therefore, the in situ TBET is a practical method for measuring the survival percentage of cells attached tightly to the culture substrate.According to the difficulty level of enzymatically detaching cells from culture surfaces and the number of spontaneously detached cells in culture supernatant, the in situ TBET or/and the hemacytometer-based one can be selected to determine the overall survival percentage of cultured adherent cells.关键词
台盼蓝染色/贴壁培养细胞/活细胞率/细胞计数Key words
trypan blue staining/ cultured adherent cell/ percent cell survival/ cell count/分类
生物科学引用本文复制引用
彭彬,吴晶晶,李烨,徐倩,唐琳,汪保和..贴壁培养细胞台盼蓝拒染试验的方法学探讨[J].激光生物学报,2011,20(2):269-273,5.基金项目
湖南省教育厅项目(N0.07A036) （N0.07A036）
