change medium of HepG2 - (Aug/16/2007 )

Hi everybody,

I work with HepG2 in RPMI 1640, 10% FCS, 2mM Glutamine. I need to change the medium of this cells from RPMI to DMEM. I'd tried already 3 times to change it and finally all the cells died!!!
To adapt the cells to the new medium, I put 75% RPMI-25%DMEM the 1 week, 50-50 the 2nd week, 25-75 the 3th week and 100% DMEM the 4th week.

Somebody can help me???????,

Hi,

it seems to be necessary to change media, otherwise I would just leave them in RPMI if they grow fine.
What you could try is to thaw cells directly into DMEM media.
I suppose you did that already, but in case not - compare basic things between the media : L-glut content, pyruvat...a.s.o.
Is the DMEM media turning pink in your incubators? Might be that it needs some adjustment of carbonate then.

Good luck

I have been growing these cells in DMEM with 10% FBS for some time. Cells seem to be quite happy. I do have to use twice strong Trypsin-EDAT to get a nice suspension, or they will form clumps. Other than that, I have no problem.