Targets in Cell Proliferation, Viability, and Cytotoxicity
Various assays are used for assessing cell proliferation, viability, and cytotoxicity. Examples include optical counting, [3H] thymidine, MTT, WST, lactate dehydrogenase (LDH) assay, and more. This selection guide aims to provide you with information that will allow you to pick the method most suitable for your specific experimental needs.
Proliferation
Measure cell count and/or DNA content to reveal cell division and DNA replication, respectively.
Methods:
– Optical counting with hemocytometer
– DNA quantification using fluorophores
Viability
Measure parameters of cellular health to determine the intracellular impact of cytotoxicity.
Methods:
– Measuring evidence of cellular metabolic activity such as esterase activity, ATP, NADH dehydrogenase activity via MTT or WST, and so on
Cytotoxicity
Measure indicators of cell death to evaluate cytotoxicity.
Methods:
– Measure the activity of lactate dehydrogenase leaked out of damaged cell membranes
– Use membrane-impermeable dyes to selectively stain inside cells with damaged membranes
Guide: Reagents Related to Cell Proliferation/Viability/Cytotoxicity Assays
The MTT and WST techniques are widely-used methods for evaluating cell proliferation and cytotoxicity. They are valued for the simplicity of their respective procedures, their safety, and the reproducibility of their results. WST (water-soluble tetrazolium) was developed by DOJINDO Laboratories in the 1990’s and is now used worldwide in cell proliferation assays. Cell Counting Kit-8 (CCK-8) is a commercial product that utilizes the WST method with improved sensitivity. Cell Counting Kit-8 is the original WST product, with which more than 5,000 papers have been published worldwide.
In order to verify the accuracy of results in a cytotoxicity assay, individual samples are measured according to multiple different principles. Cell Counting Kit-8 (code: CK04) and Cytotoxicity LDH Assay Kit-WST (code: CK12) measure dehydrogenase activity and the released amount of LDH as reflective of living and dead cells, respectively. As shown in the example below, separate measurements indicate that the number of living cells decrease, and the number of dead cells increase as a toxicant’s concentration increases.
Please watch this video for comparison between MTT and Cell Counting Kit-8.
