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Cancer Pharmacology: Provide recommendations in a multi-million dollar project
About Cancer Pharmacology: Provide recommendations in a multi-million dollar project Virtual Lab Simulation
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?
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Techniques In Lab
- Cell counting
- Drug treatment of cancer cells
- MTT assay
- EC50 determination
Learning Objectives
At the end of this simulation, you will be able to…
- Understand the principles and limitations of chemosensitivity testing in the context of in vitro based anticancer drug screening
- Conduct, analyze and interpret in vitro chemosensitivity tests using the MTT assay
- Evaluate the advantages and disadvantages of chemosensitivity tests for in vitro based screening programs
- Prepare cell cultures for chemosensitivity testing
- Determine viable cell numbers using an automated cell counter
- Conduct the MTT assay to assess cell survival following drug exposure
- Generate dose response curves and determine half-maximal effective concentration (EC50) values
- Interpret the results obtained in the chemosensitivity assay
- Understand the mechanisms of action of cyclophosphamide and epirubicin
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