I recently started culturing THP-1 cells. I know that these cells are normally cultured in RPMI 1640 media however, I need to switch them over to DMEM since I will be preparing conditioned media from them to use on some of my other cell lines (and I will also be doing it vice versa). I've been doing this gradually, by increasing the amount of DMEM and decreasing the RPMI in their media with each passage. But I've noticed that during the process the cells start forming clumps, and when they reach almost 100% DMEM, there are quite a large number of medium-large size clumps. I have also tried supplementing the DMEM with beta-mercaptoethanol, but so far it doesn't seem to be making any difference. I have cells growing in RPMI as well, and they look perfect. The FBS concentration is 10%. I've been splitting them at a constant ratio (5x, every 3rd day) and I've made sure on several occasions that they don't reach 1x10^6 cells per ml. I have seen a few papers stating that they cultured THP-1 cells in DMEM. Does anyone have any suggestions to prevent the clumping? Thank you!
hi
I do not think there is a problem . you can test thawing the cells by DMEM 10% (not gradually )
Of course, changing the culture medium may affect the expression of cell antigens.
good luck
We used DMEM almost exclusively previously, and the THP-1 cell do clump during expansion and that's never a problem. I never worked out why they clumped.
Hi,
THP-1 is generally grown in nutrient rich RPMI media. for the stress response experiments , i had to switch them to DMEM media, wherein i encountered lots of cell death . Hence i had to discontinue the propagation in DMEM. However shorter stress in DMEM ( 3 hours) was used in my experiments which did not affect the cell viability. Further DMEM comes with Low and high glucose. You can try culturing them with high glucose and assess the clumping as well as viability
Thank you all!
Dinesh Kumar Parandhaman , I am using high glucose DMEM at the moment, since we use that for our cancer cells. But, I've noticed now that when I placed RPMI cultured cells straight into DMEM media after splitting (which will have a final concentration of 20% RPMI), they look fine for the first 24 hrs, and then clumps start to appear at 48hr. So I might also try to keep my incubations to max 24 hrs then.
- Badges
- Science topic
- Similar topics
- Biological Science
- Immunology