I am working with HL-60 (suspension cell) and HuH7 (adherent cell). Normally it is seen that the cells become confluent within 3 days after subculture; while sometimes it does not. It may take 1 or 2 days more. In that case is it necessary to change the media before next subculture, as the cells are in the same media for 4 to 5 days? Is there any change in media properties like nutrition or pH?
Specially I am talking about the adherent cell where DMEM (high glucose) with 10% FBS, L-glut. and antibiotics are used. How many days the media can be good for the cells during incubation after subculture until desired confluency?
Please share your valuable experience, suggestion and any reference paper (if any).
Regards
- Make sure to count the cells before you seed them and find out the suitable cell number to be used. Sometimes it also depends on the batch of the cell used and some batches may be more stressed out than others and takes them longer time to recover from their shock especially during their first passage after taking them from liquid nitrogen,
I agree with Steve, you should always count your cells before you seed them and see the suitable cell number at each cell's data sheet.
I don't recommend you leave the cells for 5 days with the same media. You can take out some of the old media and complete with a new media, without change it all...
If the cells are not adherent, you can take some of the old media, put in a tube, centrifuge, discard the old media, ressuspend the cells and put them back at the flask.
And answering your question, with time cells consume the nutrients of your media, and the pH change is an indication of this. Some cell media come with phenol red, a pH indicator.
Best of luck!
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