This video describes the colony formation assay, which helps determine lung cancer cells' ability to proliferate indefinitely and form individual colonies. We can analyze the effect of an anticancer agent on the colony-forming cells by analyzing the changes in the number and/or size of the resulting colonies.
Protocol
1. Colony Formation Assays Culture A549 cells at a concentration of 20,000 cells/cm2 in 15 mL of RPMI1640 cell culture medium supplemented with 10% FBS (Fetal Bovine Serum) and 1% antibiotics in a 75 cm2 flask in a CO2 incubator at 37 °C and 5% CO2. On the day of the experiment, determine the cell concentration by using a hemocytometer. Collect 50,000 cells by centrifugation at 400 x g for 7 min in a 1.5 mL microcentrifuge tube and re-suspend c…
Colony Formation Assay: Assessing the Efficacy of Anticancer Agents on Colony-Forming Lung Cancer Cells. J. Vis. Exp. (Pending Publication), e20243, doi: (2023).