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Conflicting opinions - To change media or not to change media (May/30/2006 )

I am just wondering ... I have been told that I shouldn't change media on my cells that it is a better idea to split them then to change their media. I was told that changing the media would make the cells grow too fast (but wouldn't splitting do the same thing) I feel like I should change the media at least every few days, especially on some cell lines. I work primarily with hepatoma cell lines, though my previous experience was with lymphocytes (so perhaps that is what is clouding my judgement). I have one cell line that tends to shed a lot of viable cells into the media. It gets so bad that at times that it is hard to see the adherent cells through the cells in suspension. I was told though to let this cell line grow and not to touch it for about two weeks (no changing media/no splitting) and then split it. But I have a strong desire to change the media when there are that many cells floating in suspension. I'm just curious what others think of this. What would you guys do?


I split my cells (different 293s ) as they get confluent, which is when the medium is also orange-yellowish.


it's different based on cell type and other factors...I would contact the vendor (or, if you are isolating primaries from tissue, a vendor) of your cell type and see what is recommended


Dear Jamie,

if you have been advised the let the cells sit ... let them be.

If you change the media (remove the floaties), you are disrupting the environment and placing stress on the cells. Taking the flask in and out of the incubator also places stress and heat-shock on the cells.

Is the media turning orange-yellow during the two week culture period??


The media does tend to change color during the two week period (I am fairly certain this is due to how many cells are floating in suspension). I don't think the cells are doing poorly though. The literature I received from ATCC with the cell line suggests media renewal two to three times a week. It also says that it is normal for the cell line to shed viable cells into the media and that it can make the media appear turbid.

I feel a bit better now though leaving the cells alone.

I appreciate all of the responses.