I just started culturing MCF7 cells but they are growing extremely slowly, they do not look healthy, and there are a lot of floating cells. I am growing them in a 10cm cell culture dish with · RPMI-1640 (2mM L-glutamine, 10mM HEPES, 1mM sodium pyruvate, 4500 mg.L glucose, and 1500 mg/L sodium bicarbonate) supplemented with 10% FBS and 1% pen/strep. I have also started culturing one dish in DMEM supplemented with 10% FBS, 1% NEAA, and 1% pen/strep. Both appear to look the same. Can anyone explain why my cells look like this?
Dear Allison,
to me it looks like that the cells were not able to adhere to the surface of the dish. Are you sure you are using (commercial) dishes treated for tissue culturing? (These cells will not grow on untreated Petri dishes.) If so, try another charge (lot) or supplier.
Also, when using DMEM, MCF-7 cells do not need further supplementation with non-essential amino acids.
Hope this will help you and the cells.
Angela
Hi Allison,
Indeed as you mention, cells are not really attaching so it can be an issue as Angela has mentioned, with untreated dishes. Regarding media, I don't think media is a factor but in any case you can confirm in the datasheet of the supplier which media suplementation is best and at what concentration you should be seeding. Also, other factors that can interfere is presence of DMSO from freezing media, contamination, insufficient CO2, or they were poorly stored and are dying out which would be my second guess (-80ºC for too long instead of liquid nitrogen). You can try to pass them to a T25 and see if they attach and recover or otherwise, best to get from a new batch.
Thank you Angela and Sofia! I am using treated cell culture plates however I can try coating them with collagen to see if that helps the cells attach better. If this doesn't work maybe I will reach out to ATCC.
- Badges
- Science topic
- Similar topics
- Biological Science
- Molecular Cell Biology
- Culture Cells
- Cell Line