Cytotoxicity Test

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Cytotoxicity test
Cell cytotoxicity is a simple cell killing event caused by cells or chemicals and does not depend on the apoptotic or necrotic mechanism of cell death.Sometimes cytotoxicity testing is required for specific substances, such as drug screening.

Technical details
Normal cells are metabolically vigorous and their mitochondria contain succinate dehydrogenase, which can reduce tetrazolium salts (e.g., MTT, XTT, WST-1, and WST-8, etc.) to purple crystalline substances that deposit around the cell. Then we can use a microplate reader to read the OD value (optical density), thus detecting the status of cell proliferation.
Cell counting kit-8 (also known as CCK-8) is a rapid and highly sensitive assay based on WST-8 (Water Soluble Tetrazolium-8) and widely used for cell cytotoxicity analysis.
The rationale is: WST-8 [chemical name: 2-(2-Methoxy-4-nitrophenyl)-3-(4-nitrophenyl)-5-(2,4-disulfophenyl)-2H-tetrazolium Sodium Salt] is reduced by dehydrogenases in cells under the reaction of the electron carrier 1-methoxy-5-methylphenazinium sulfate dimethyl ester (1-methoxy PMS) to a highly water-soluble yellow formazan product (formazan dye).The number of formazans generated directly correlates to the number of living cells.This property can therefore be exploited to directly carry out cell cytotoxicity assays.The more rapidly the cells proliferate and the more cells proliferate, the darker the color is, and the more toxic the cells are, the lighter the color is.For the same cells, there is a linear relationship between the intensity of the color and the number of cells.


Case study


Reference
Liu, Qing-Hua et al. “Reduced expression of annexin A1 promotes gemcitabine and 5-fluorouracil drug resistance of human pancreatic cancer.” Investigational new drugs vol. 38,2 (2020): 350-359. doi:10.1007/s10637-019-00785-5


Service turnaround and deliverables

Service turnaround

5~10 business days, depending on cell growth and experimental design

Customer provides

cell lines, drugs, drug treatments and experimental conditions parameter settings

Deliverables

raw data, analytical results, experimental reports, remaining drugs

Other methods to detect cytotoxicity
Principle Features
ATP assay The fluorescence of luciferase and luciferin under the action of ATP can be detected/quantified by chemiluminescence instrument Suitable for detection of reproductive capacity, growth inhibition and death of cells after being stimulated by drugs, poisons and biological factors
LDH assay To detect the LDH activity in cell culture thus detecting cytotoxicity Simple and rapid; suitable for detection of cytotoxicity caused by drugs or radiation