I have a colony formation assay. I am seeding cells in a 96-well plate. The cell line is NTERA, 150 cells per well. I am seeding the cells with DMEM fortified with 10% FBS. Each well will have 200uL of this media. I will be incubating the treated cells for 14 days. I will be counting colonies on days 4, 7, 10, and 14.
How often should I remove the old media and replenish the wells with fresh media?
Thanks in advance.
In a colony formation assay, the frequency of media replenishment depends on the growth rate of the cells and the duration of the assay. For your specific experiment with NTERA cells, which have a doubling time of approximately 30 hours, replenishing the media every 3-4 days should be sufficient to support cell growth and colony formation.
NTERA doubling time is actually 50-60 hours. Should I still go with 3-4days?
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