The WST survival assay: an easy and reliable method to screen radiation-sensitive individuals

Abstract

An easy, fast and reliable method was developed to screen hundreds of Epstein-Barr virus-transformed cell lines (lymphoblastoid cell lines, LCLs) for radiation sensitivity that were generated from lymphocytes isolated from young lung cancer patients. The WST-1 test explores the metabolic activity of the mitochondria as an indicator for the vital status of cells. Cell proliferation as well as indirect cell death can be quantified by this method on a large scale in microtiter plates. Cell survival was measured at 24- and 48-h post-irradiation with 10 Gy ((137)Cs source) by the WST-1 assay and Trypan blue staining. To set up the experimental screening conditions and to establish a positive and a negative control, an ATM-mutated cell line from a radiation-sensitive ATM patient and an ATM proficient cell line from a healthy brother were compared. An optimal differentiation between the two cell lines was demonstrated for 10 Gy and 24- and 48-h cell growth after irradiation. Upon screening 120 LCLs of young lung cancer patients under these conditions, 5 of them were found to be radiation sensitive to a high degree of statistical significance. The results have been confirmed by a second laboratory by means of Trypan blue testing. The WST-1 test represents an efficient and reliable method by means of screening for radiation-sensitive cell lines.

Comprehensive Profiling of Radiosensitive Human Cell Lines with DNA Damage Response Assays Identifies the Neutral Comet Assay as a Potential Surrogate for Clonogenic Survival

“…6B, Table 1). These data suggest a fundamental relationship between abnormal mitochondrial respiration and radiosensitivity and are consistent with previously published reports (32, 36). …”

“…Mitochondrial respiration rates were reduced in the majority of radiosensitive LCLs tested (68%) (Table 1). A recent report using LCLs generated from young lung cancer patients explored the metabolic activity of mitochondria as an indicator of cell survival as a possible diagnostic test for radiosensitivity (36). In addition, Guo et al .…”

The inter-individual variability outperforms the intra-individual variability of differentially expressed proteins prior and post irradiation in lymphoblastoid cell lines

“…GM03189 is commercially available (Coriell Institute, NJ, USA) and represents a radiosensitive LCL of an AT-patient. The LCLs 4060-200 (radio-sensitive) and 20037-200 (normal) are EBV immortalized B-lymphocytes from patients of the nationwide, multicentre LUCY-study (www.helmholtz-muenchen/epi/) and their individual radiation response was characterized (Guertler et al, 2009;Maroschik et al, 2014). All cell lines were grown at 37 C in a humified atmosphere of 5% CO 2 .…”

“…The other two cell lines derived from lung cancer patients of the LUCY study (Rosenberger et al, 2011). The LCL 4060-200 shows a radio-sensitive phenotype, whereas the second LUCY-cell line 20037-200 reacts normal or radioresistant in contrast to the other cell lines in the different endpoints that were tested (Guertler et al, 2009;Maroschik et al, 2014).…”

Identification of Lung Cancer Related Function Modules Based on Co-Expression Network

“…联：Nomori H.和 Mori T.等人 [38] 经过四年，对 212 个 肺癌患者的跟踪研究发现， HTLV-I 病毒可能是与验证 或者免疫反应有关的细支气管肺泡癌的致病风险。此 外，对于模块 351 中包含的基因 SMAD5 和 PPCS，现 有研究 [39,40] 发现它们是与 Epstein-Barr(EB)病毒相关 的，而 Guertler A.等人 [41] 又发现 EB 病毒可以从年轻的 肺癌患者身上发现放射性敏感性。而模块中的另一个 基因 FUT6，Norden R.等人 [42] 发现疱疹病毒可以通过 基于共表达网络挖掘肺癌相关模块 Copyright © 2013 Hanspub …”