Clonogenic Survival Assay Protocol
The first clinical trial of a chemotherapeutic agent for the treatment of cancer took place in 1946 with limited success. While doctors searched for new treatments, researchers developed the Clonogenic Cell Survival Assay to test these new agents before testing them in patients.
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Description
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The clonogenic survival assay measures the toxicity of a treatment on cells at a wide range of doses. The type of cell measured depends on the assay, such as cancer cells for toxicity or normal cells for side effects. The assay readout specifies the cell ability to reproduce and form clones.
Clonogenic
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In its basic format, the treated cells are diluted and grown on petri dishes. If a cell is viable, it divides and over time forms a small mound of cells called clones. All the cells are genetically identical. At the end of the experiment, the clones are stained and counted to determine survival rates at each dose.
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Modifications
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Modern assays can be done in different formats. For example, flow cytometers use laser beams to deposit one treated cell in each well of a 96-well plate. The scientists can use reagents such as the alamar blue assay, which measures metabolic activity, to find and quantify the clones.
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- Photo Credit culture plates and flasks image by Fotocie from Fotolia.com
