A Simple Protocol for Using a LDH-Based Cytotoxicity Assay to Assess the Effects of Death and Growth Inhibition at the Same Time

Abstract

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.

Interlaboratory Evaluation of in Vitro Cytotoxicity and Inflammatory Responses to Engineered Nanomaterials: The NIEHS Nano GO Consortium

“…In contrast, the CPPs conducted phase II studies using protocols developed after identifying and solving technical problems in phase I. The CPPs selected MTS and LDH assays because they are the most commonly used single end point cell viability assays for nanotoxicity studies in the literature (Smith et al 2011; Wang et al 2010; Xia et al 2008a). The relative simplicity of the assay procedures allowed the studies to be conducted concurrently.…”

Protective Effects of Berry Extracts on Hydrogen Peroxide-induced Rat Brain Neuronal Cell Damage in vitro

“…Release of LDH (indicating loss of plasma membrane integrity [13]) was measured using an LDH detection kit according to the manufacturer's instructions (Sigma, MO, USA). PC-12 cells were pelleted by centrifugation (250 × g) for 5 min at RT, 100 μL of the supernatants were transferred into new wells, and LDH was determined using the in vitro toxicology assay kit.…”

<i>In vitro</i> oral epithelium cytotoxicity and <i>in vivo</i> inflammatory inducing effects of anesthetic rice gel

“…In fluorescent images of the epithelial cells staining with DAPI and F-actin, the nucleus and cytoplasm, respectively, of the cells were obviously seen confirming the cells could be survived after contact with lidocaine and prilocaine. For investigation of cytotoxicity using LDH assay, high LDH release indicates high amount of cell death (29). The results of LDH assay revealed that lidocaine and prilocaine at 5-20% had no toxic effect to the cells.…”