Cellular Bioluminescence Based Assay: A High Throughput Method for Combinatorial Drug Screening
Transcript
Coat 96-well optical bottom plates with an extracellular matrix mixture such as Matrigel and incubate them for 1 hour at 37 degrees Celsius. Remove the excess mixture and gently rinse the plates once with PBS.
Then, seed XG387-Luc cells at a density of 1,000 cells in 100 microliters of culture medium into each well of the coated plates and culture them overnight. On the following day, observe the cells under a microscope to confirm their attachment to the plate.
Next, prepare a 200-micromolar temozolomide solution and 2-micromolar solutions of the targeted agents in culture medium. For combination drug screening, remove the blank medium and add the medium containing either 200-micromolar temozolomide or 2-micromolar targeted agent, or a combination of both, into each well for 3 technical replicates per treatment.
Incubate the plate at 37 degrees Celsius and 5% carbon dioxide for 3 days. Take images of the cellular bioluminescence in the plate using the IVIS spectrum imaging system. Using the built-in software, create multiple circular areas of the region of interest and quantify the cellular bioluminescence.
