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How often should you change your cell culture media?

In general, media should be changed every 2-3 days. However, this will depend on the cell culture type cell density and volume of medium used per unit of surface. Allowing the medium to turn yellow as a consequence of the acidic pH generated by cell metabolic activity should be avoided as it can lead to cell death and phenotypic changes in the culture. Moreover not all the media will turn yellow before being exhausted of nutrients.

Usually (though not always, depending on the cell culture type), the timings of when you change your cell culture media coincide with what is called passaging or split. In general, this involves the transfer of either adherent or suspension cell cultures once they have grown to confluency, to ensure continued cell culture growth:

  • Adherent cells are usually passaged by treating the cell culture with an enzyme that helps to lift adherent cells from the surface, before a fraction of this suspension is reseeded to a clean, unused plate/flask and additional media is added. The plate/flask manufacturer usually has information on the media volume for each dish/flask for proper gas exchange. Cells should be seeded at a density that is most optimal for proliferation for the specific cell type and should form a monolayer.

  • In suspended cells, a proportion of the cells suspended in the cell culture medium are removed from the container and placed into a fresh flask/dish to maintain the correct seeding density for that particular cell type in a given flask/dish.


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