In this simulation, you will learn which tools are needed to screen thousands of chemical compounds in order to identify promising candidates for the treatment of different types of cancer. In particular, you will gain insight into the principles and limitations of widely used in vitro assays in the context of anti-cancer drug screenings. As a pharmacology expert, you will test the robustness of those commonly used techniques and help a screening facility to assess the risk for false negative results.

Prepare cells and drugs for chemosensitivity testing

Your main task in the Cancer Pharmacology simulation is to reproduce the screening facility’s chemosensitivity in vitro protocol to investigate the cause of false negative results. The protocol in question is a widely used cell survival assay using MTT reagent and performed with conventional cancer cell lines. The Project Leader suggests to perform the test with two well-known chemotherapy drugs: epirubicin and cyclophosphamide. In this part of the simulation, you will learn how to perform common steps of chemosensitivity testing and learn about basic pharmacology tools. Most importantly, you will count living cells of a cell culture solution and calculate how much of the culture you need for your experiment. You will also prepare stock solutions of both drugs with a given concentration to create a dilution series for the treatment of the cancer cells.

Perform the MTT assay

Once the cells have been prepared and incubated with the drugs, it is time to use MTT as a cell survival indicator. You will be able to follow on a molecular level how MTT specifically reacts with mitochondrial enzymes leading to a color colorimetric change that can be measured with a spectrophotometer. In this mission, you will work with a multi-channel pipette and 96 well plates to perform the assay as used in many pharmacology labs worldwide.

Determine EC50 and interpret your results

The result of the serial drug dilution in the MTT assay will provide the basis for creating a dose-response curve and for determining the half-maximal effective concentration (EC50) of both clinically used drugs. Which recommendations will you provide the Project Leader with about the usefulness of the commonly applied in vitro assays?