A simple protocol for using a LDH-based cytotoxicity assay to assess the effects of death and growth inhibition at the same time

• Published in: PloS one Vol. 6; no. 11; p. e26908
• Main Authors: Smith, Shilo M, Wunder, Michael B, Norris, David A, Shellman, Yiqun G
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 17.11.2011; Public Library of Science (PLoS)
• Subjects: Analysis; Apoptosis; Assaying; Biological Assay - methods; Biological research; Biology; Cancer; Cell cycle; Cell Death; Cell growth; Cell Line, Tumor; Cell proliferation; Cell Proliferation - drug effects; Cell Survival - drug effects; Cytotoxicity; Dehydrogenases; Dermatology; Dose-Response Relationship, Drug; Drug dosages; Growth; High-throughput screening; Humans; Indoles - pharmacology; Inhibition; Kinases; L-Lactate Dehydrogenase - metabolism; Medical research; Medicine; Metabolism; Mortality; Pharmaceutical sciences; Protocol; R&D; Reproducibility of Results; Research & development; Screening; Sulfonamides - pharmacology; Toxicity; United States > US; Colorado
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0026908

Analyzing the effects on cell growth inhibition and/or cell death has been an important component of biological research. The MTS assay and LDH-based cytotoxicity assays are two of the most commonly used methods for this purpose. However, data here showed that MTS cell proliferation assay could not distinguish the effects of cell death or cell growth inhibition. In addition, the original LDH-based cytotoxicity protocol grossly underestimated the proportion of dead cells in conditions with growth inhibition. To overcome the limitation, we present here a simple modified LDH-based cytotoxicity protocol by adding additional condition-specific controls. This modified protocol thus can provide more accurate measurement of killing effects in addition to the measurement of overall effects, especially in conditions with growth inhibition. In summary, we present here a simple, modified cytotoxicity assay, which can determine the overall effects, percentage of cell killing and growth inhibition in one 96-well based assay. This is a viable option for primary screening for many laboratories, and could be adapted for high throughput screening.