Training | Drug Treatment
ADRI Training Video - Drug Treatment
1. Current drug-based therapies for asbestos-related cancers are suboptimal and drug resistance and toxicity issues limit their clinical efficacy. Therefore there is a need to develop and test novel drugs in a laboratory setting before they can undergo clinical trials.
2. Drug treatments are routinely carried out on the various asbestos-related cancer cell lines at ADRI. The aim is to elicit a cellular response that will enable us to predict their therapeutic potential for the treatment of asbestos-related cancers.
3. Following trypsinization of the cells, the cell suspension is transferred to a centrifuge tube and subjected to centrifugation to pellet the cells. The resulting cell pellet is then resuspended in a small volume of growth media.
4. A small aliquot of the cell suspension is combined with a trypan blue stain, mixed and loaded onto a cell counting slide.
5. The cell counting slide is inserted into an automated cell counter, which counts the number of cells on the slide and calculates the number of cells per mL of the original cell suspension.
6. Taking into account the known concentration of cells in the cell suspension, the cells can be diluted further with growth media to obtain a desired concentration.
7. Equal volumes of the diluted cell suspension are then seeded within each well of the cell culture plate and the cell culture plate is then placed into the cell incubator to allow the cells to adhere and grow.
8. The next day the cells are treated with the drug of interest at varying doses and the cells are then placed back into the cell incubator.
9. After a 72hr drug exposure period, the growth media is removed from each well and a fluorescent dye is added to stain the viable cells. The cells are left to react with dye for 4hrs. The viable cells are able to react with the dye and produce a fluorescent signal, which is quantified upon inserting the plate into a spectrophotometer.
10. The number of viable cells across the varying drug concentrations are graphically plotted in the form of a dose response curve using graphing software. The half-maximal inhibitory concentration of the drug, known as the IC50, can then be interpolated from the graph. This provides us with an indication of how effective or ineffective the drug is at inhibiting tumor cell growth.
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