Fraunhofer-Verbund Life Sciences
Figure 1: Cell viability after application of 2 compounds (A and B) compared to the application of a small molecule displaying cellular toxicity in Hep-G2 cells.
Determination of cell viability after application of small molecules in vitro
Species
Adherent cell lines
Field of application:
This assay is used to test the viability of cells after incubation with small molecules and to determine the toxic effect of respective substances. Damaged cells release the intracellular enzyme lactate dehydrogenase which can be determined using a fluorometric assay. The amount of enzyme activity correlates with the degree of toxicity of the substances.
Three different cell lines are mainly used:
- Intestinal cell line Caco-2 as relevant cell model for the intestinal transport after oral intake
- Hepatic cell line HepG2 and neuronal cell line SH-SY5Y as relevant cell models for the in vivo pathway of drugs
- > 100 other permanent cell lines available upon request
This model can be used for the following fields of application:
- Pre-screening for toxic substrates in vitro
Endpoints/Outcome parameter
Percentage of the viability of cells after substance incubation compared to untreated control cells.
Readout parameter
Fluorometric measurement of lactate dehydrogenase activity.
Quality management and validation
- For each test series a control compound with known toxicity is used
- Internal quality management
- Assay can be performed according to GLP (certification within category 9)