Cellular Bioluminescence Based Assay: A High Throughput Method for Combinatorial Drug Screening
Transcript
Cellular bioluminescence is a highly sensitive and rapid approach for large-scale screening of anti-cancer drugs.
This technique uses tumor cells expressing luciferase – an enzyme that catalyzes a light-producing chemical reaction, causing the host cells to glow in the dark. The intensity of the emitted luminescence is directly proportional to the live cell density.
To begin, seed luciferase-expressing cells in an optically clear culture plate coated with an extracellular matrix or ECM and incubate.
During incubation, the ECM promotes the attachment of cells to the culture well.
Next, take concentrated stocks of the drug combination and add different volumes of culture media to prepare serial dilution of the drug mixture.
Now, remove the culture medium from the adhered cells. Add different dilutions of the drug combination and incubate.
During incubation, the drug combination acts synergistically on cellular targets.
Now, remove the drug-containing medium and add fresh medium containing luciferin.
Luciferin diffuses across the membrane and enters the cells. The luciferases expressed by the cells use cellular ATP to oxidize luciferin and emit light signals.
Quantify the cellular bioluminescence under an imaging system.
A decrease in light emission with increased drug concentration indicates reduced cell viability. This correlates to the drug's antiproliferative effect on the target cells.
