Measurement of cytotoxicity by ATP-based luminescence assay in primary cell cultures and cell lines
Measurement of cytotoxicity by ATP-based luminescence assay in primary cell cultures and cell lines
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DOI：
10.1016/s0887-2333(97)00060-x
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发表时间：
1997-10-01
影响因子：
3.2
通讯作者：
Andreotti, PE
中科院分区：
文献类型：
--
作者：
Cree, IA;Andreotti, PE
药物发现和毒理学安全性测试都需要可靠的体外细胞毒性测试。理想情况下，这样的测试应该是客观的，定量的，可重复的，并能够自动化。许多试验很好地满足了这些标准，但最近已经清楚的是，测试细胞的分子表型和不同细胞类型之间的复杂相互作用可以从根本上改变对单个试剂的反应。原代细胞培养物和细胞系之间观察到的差异使得最好使用原代培养物来评估毒性，但使用原代细胞培养物的问题相当大，因为可用于测试的细胞数量通常很少。最近，我们已经开发了一种短期的细胞培养试验的基础上检测ATP的荧光素酶反应。四种药物/试剂可以在一个96孔微孔板中以7种稀释度一式三份进行测试，对于细胞系，1000个细胞/孔，或对于原发性肿瘤组织，10，000个细胞/孔。所需的细胞数量少是这种方法的主要优点。最初作为肿瘤化疗敏感性试验开发，该试验已显示出相当大的前景，作为一个通用的体外毒性试验，允许细胞系和原代组织培养物进行测试。在良性组织活检以及肿瘤中存在敏感性的异质性。细胞内的分子改变和不同细胞类型的相互作用已经在许多不同的模型系统中使用该测定法得到解决，这表明该技术可能具有更广泛的应用。(C)1997 Elsevier Science Ltd.
Drug discovery and toxicological safety testing share a need for dependable in vitro cellular toxicity tests. Ideally such tests should be objective, quantitative, reproducible and able to lend themselves to automation. A number of assays fulfil these criteria well, but recently it has become clear that the molecular phenotype of the cell tested and the complex interplay between different cell types can radically alter the response to individual agents. The differences observed between primary cell cultures and cell lines make it preferable to use primary cultures for assessment of toxicity, yet the problems of using primary cell cultures are considerable as the number of cells available for testing is often small. Recently, we have developed a short-term cell culture assay based on the detection of ATP by the luciferin-luciferase reaction. Four drugs/agents can be tested in triplicate at seven dilutions in one 96-well microplate with 1000 cells/well in the case of cell lines, or 10,000 cells/well for primary tumour tissue. The small number of cells required is a major advantage of this method. Initially developed as a tumour chemosensitivity assay, the assay has shown considerable promise as a general in vitro toxicity assay allowing both cell lines and primary tissue cultures to be tested. Heterogeneity of sensitivity is present in benign tissue biopsies as well as tumours. Molecular alterations within the cell and the interplay of different cell types have been addressed in a number of different model systems using the assay, suggesting that this technology may have more general application. (C) 1997 Elsevier Science Ltd.