THE USE OF ATP BIOLUMINESCENCE AS A MEASURE OF CELL-PROLIFERATION AND CYTOTOXICITY
THE USE OF ATP BIOLUMINESCENCE AS A MEASURE OF CELL-PROLIFERATION AND CYTOTOXICITY
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DOI：
10.1016/0022-1759(93)90011-u
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发表时间：
1993-03-15
影响因子：
2.2
通讯作者：
FLETCHER, J
中科院分区：
文献类型：
--
作者：
CROUCH, SPM;KOZLOWSKI, R;FLETCHER, J
用三磷酸腺苷(ATP)生物发光法确定培养细胞数与荧光素-荧光素酶反应测得的发光值之间是否存在线性关系。经Spearman等级相关系数(p>；0.00001)检验，外周血单个核细胞、MOLT-4、HL-60、TF-1、NFS-60和L-929细胞系之间存在显著的相关性。然后，这些观察结果被用来确定ATP生物发光是否可以作为衡量细胞增殖的合适的替代氚胸苷摄取的指标。细胞系MOLT-4、HL-60、TF-1和NFS-60显示胸腺嘧啶核苷摄取与三磷酸腺苷生物发光之间有很强的相关性(p>；所有类型的细胞均为0.00001)。此外，该方法还能检测到粒细胞巨噬细胞集落刺激因子(GM-CSF)和粒细胞集落刺激因子(G-CSF)对TF-1和NFS-60细胞的细胞因子依赖性增殖作用。该方法还能准确检测肿瘤坏死因子对L细胞的细胞毒作用。因此，在许多不同的生物检测中使用ATP生物发光来检测细胞因子的活性似乎是可能的。
Adenosine triphosphate (ATP) bioluminescence was used to determine whether there was a linear relationship between cultured cell number and measured luminescence using the luciferin-luciferase reaction. In all the cells tested including peripheral blood mononuclear cells (MNC), MOLT-4, HL-60, TF-1, NFS-60 and L-929 cell lines there was a significant correlation as determined by Spearman's rank correlation coefficient (p > 0.00001). These observations were then used to determine whether ATP bioluminescence could be used as a suitable substitute for tritiated thymidine uptake as a measure of cell proliferation. The cell lines MOLT-4, HL-60, TF-1 and NFS-60 showed a strong correlation between thymidine uptake and ATP bioluminescence (p > 0.00001 for all cell types). Additionally the ATP method could detect the cytokine dependent proliferation on TF-1 and NFS-60 cells by granulocyte-macrophage colony-stimulating factor (GM-CSF) and granulocyte colony-stimulating factor (G-CSF) respectively. The tumour necrosis factor alpha (TNF)-induced cytotoxic effect on L-929 cells could also be accurately detected using this method. It would therefore appear to be possible to use ATP bioluminescence in the detection of cytokine activity in a number of different bioassays.