I work with Hella in RPMI 1640. I need to change the medium of this cells from RPMI to DMEM. How to change these cell lines to adapt the cells to the new medium?
There is not big difference in the composition of both medium RPMI1640 and DMEM. By lowering the FBS concentration (between 3-4%) in DMEM medium the Hella cells can adapt in better way without morphological changes. However, Hella cell line is itself a very tough cell line can adapt any cell culture medium very easily but experimental results could be not similar after change of medium.
RPMI to DMEM should take no adaptation at all. Go straight from medium to the next. We commonly go from DMEM in flasks to JMEM in suspension. JMEM is similar to RPMI.
I already tried to change the media for HeLa cell from DMEM to RPMI without any adaptation and I succeed. Just go straightly no need to add or reduce anything at all.
Which cell line is it? Some cells can grow in both RPMI and DMEM. Some not. Why do you have to adapt the cells? For short incubation 2-3h there should be no difference. For long culture you can select some clone of cells and finally your cells won't be the same.
yes, there is no not much difference in composition of rpmi and dmem, so cells can be directly get adapted in another media, but even if you find difficulty, then start with mixture of both the media like RPMI:DMEM = 90:10, 75:25,50:50,30:70,15:85,0:100 with each subculture.
I would be very careful about repeating experiments after the switch. Our laboratory, as well as others, have demonstrated significant changes in certain mRNA and protein expression upon similar switches, AMEM to DMEM was profound. When making the switch, try to look at mRNA arrays for critical genes to determine that the expression of proteins of interest are not changing. mRNA is indirect, but better than nothing. As for the adaptation, Paromita gives a great example of acclimation. Another is to just thaw a new vial and begin again in DMEM.
I agree with Gregory. pending on what you want to do you have to be extremely careful to assume that mRNA expression and behaviour will be the same after the switch. While the medias are very similar you can expect still a lot of changes in gene-expression. We published about changes using defined media vs RF10 in melanoma cells and have been surprised ourselves.
Stem Cells. 2012 Feb;30(2):336-43. doi: 10.1002/stem.786. Stem cell media culture of melanoma results in the induction of a nonrepresentative neural expression profile.
If important, you may want to do a gene-expression array before and after.
Andreas, that was an eye opening paper to me when I first read it. Not to beat a dead horse, but we published these manuscripts:
W.J. Roth, D.J. Lindley, S.M. Carl and G.T. Knipp. The Effects of Intra-Laboratory Modifications to Media Composition and Cell Source on the Expression of Pharmaceutically Relevant Transporters and Metabolizing Genes in the Caco-2 Cell Line. J. Pharm. Sci. 101(10):3962-3978 (2012; EPub: July 11, 2012).
D.J. Lindley, W.J. Roth, S.M. Carl and G.T. Knipp. The Effects of Media on Pharmaceutically Relevant Transporters in the Human HT-29 Adenocarcinoma Cell Line: Does Culture Media Need to be Controlled? J. Pharm. Sci. 101(4):1616-1630 (2012; EPub: Dec 28, 2011).
In addition, we have data for two other papers including a clonal line where the differences based on media switches were profound. I would also look up publications by Donna Volpe at the FDA on the complications associated with variability. It is just hard to compare data from studies in one media to those in another.
I hope we are being overly cautious and it works out for you. I would rather see you not have any issues because you took the proper precautions versus the alternative.
Yes, do a gradual change and if you are working on metabolic pathways then you be somewhat okay.
Best wishes
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line