Cytotoxicity
Endothelial cells and cytotoxicity
Endothelial cells line the intimal surface of blood vessels forming the interface between blood and tissue. A potential drug will primary contact endothelial cells and a cytotoxic effect on these cells will be essential in evaluation of a compound in the early drug development process.
As a service for industry and institutes we offer in vitro assays on human endothelial cells to evaluate the physiological response of a compound at the cellular level.
Direct cell viability assay
Fig. A
(Click to enlarge)
Monolayers of endothelial cells are incubated in absence (Fig. A) and in presence of the compound (Fig. B). Subsequently, viable cells are stained with the fluorescent dye calcein-AM. Cell viability is quantified by software supported image quantification. Results are expressed in relation to untreated controls.
Fig. B
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Fig. A: Human dermal microvascular endothelial cells form a typical monolayer in cultur
Fig. B: In presence of a cytotoxic compound, cell death leads to a distinct desintegration of the monolayer
MTS - assay for assessment of metabolic activity
After incubation in presence of the compound, a tetrazolium salt (MTS) is added. Viable cells are able to reduce MTS to formazan which is quantified Cytotoxicity is assessed in percent of a control (compound omitted).
LDH release assay for assessment of membrane integrity
The activity of LDH released from damaged cells is determined in absence and presence of the compound. An increased LDH activity in a culture supernatant corresponds to lysis of cells and indicates cytotoxic effects.
Cell proliferation assay (Ki67 antigen expression)
The Ki67 antigen is limited to cells which have entered the cell cycle and represents a valuable marker for cell proliferation. The effect of a compound on Ki67 expression is examined in a quantitative enzyme immuno assay.
For further information please contact: info@amplab.de