PBS wash in between media change - (May/31/2006 )
hmm...this is really a very simple question but i need to confirm. i'm really new in cell culture thing.
i've been checking protocols but i can't seem to get the answer.
do I need to wash my cells with PBS in between media change?
hi annlomeda! if you are simply changing the spent media with fresh media then there is no need to wash the cells with PBS. but if you are using your cells for special purpose like stimulation, FACS then you have to wash the cells with PBS according to the requirement
I dont wash cells if I am only changing media. But I have seen some people wash cells even if its just changing media.
I don't wash cells when changing media for suspension cells (I split them, discard 35 ml, take the remaining 5 ml and add 35 or so for a big culture flask). For adherent cells: if just changing the media: no wash necesarry, but for splitting them with trypsinisation: you need to wash as remaining serum on your cells inhibits trypsin from loosing your cells.
if you are just replenishing media on the cells (removing spent media or replacing with fresh selective media), and the cells are adherent, you need not wash the cells with PBS.
If you are replacing the old media with media containing different components i.e. refeeding with low-serum or serum-free media, or adding a new component, it is often better to wash the cells prior to the addition of the new media to remove inhibiting agents etc. You can use either PBS or the new media to wash.
If the cells are non-adherent, and you just need to replenish the media, centrifuge and resuspend in the new media. If you are changing media, centrifuge, resuspend in PBS or new media and collect by centrifuging again and then resuspend the pellet in the new media for culturing.
Hope this helps :-)
wow! i was very much enlightened! hehe thanks a lot guys!
what if you're replacing media with the same media + drug?
You may not need to wash the cells with PBS after removing the old media...just add the same media +drug.
i'm culturing monolayer and i prefer to wash it. hate to see if there are dead cells/debris... actually up to your own preference.
I wash the cell with PBS between media change when i find ther is contamination in the flask.
washing the cells with PBS removes the contamination.
all the best