Department of Pharmaceutical Technology and Biopharmaceutics, Johannes Gutenberg-University, Staudingerweg 5, 55099 Mainz, Germany.
Radiat Oncol. 2012 Jan 3;7:1. doi: 10.1186/1748-717X-7-1.
For studying proliferation and determination of survival of cancer cells after irradiation, the multiple MTT assay, based on the reduction of a yellow water soluble tetrazolium salt to a purple water insoluble formazan dye by living cells was modified from a single-point towards a proliferation assay. This assay can be performed with a large number of samples in short time using multi-well-plates, assays can be performed semi-automatically with a microplate reader. Survival, the calculated parameter in this assay, is determined mathematically. Exponential growth in both control and irradiated groups was proven as the underlying basis of the applicability of the multiple MTT assay. The equivalence to a clonogenic survival assay with its disadvantages such as time consumption was proven in two setups including plating of cells before and after irradiation. Three cell lines (A 549, LN 229 and F 98) were included in the experiment to study its principal and general applicability.
为了研究照射后癌细胞的增殖和存活情况,我们对基于活细胞将黄色水溶性四唑盐还原为紫色不溶于水的甲臜染料的 MTT 比色法进行了改良,从单点比色法改为增殖测定法。该测定法可使用多孔板在短时间内对大量样本进行检测,并且可以使用微孔板读数仪半自动进行检测。在该测定法中,通过数学计算得出存活这一参数。实验证明,对照组和照射组的指数生长是多次 MTT 测定法适用性的基础。该测定法与克隆存活测定法具有等效性,但其也存在时间消耗等缺点,这在包括照射前后细胞铺板的两种设置中得到了证明。我们在实验中纳入了三个细胞系(A549、LN229 和 F98),以研究其主要和普遍适用性。