I am working on MCF-7 cells...whenever i revive the cells they get attached and for the first two days their morphology is very good..(edges glow when u view under microscope and cells seem great just like ATCC morphology)...but on 3rd day cells start detaching and by 4th and 5th day all the cells detach and die..I have tried almost everything like reducing the trypsinization time and centrifuging speeds to reduce the damage to cells...but its not helping... Also if someone has a good stock of MCF-7 cells please send it to me. That would be very kind of you..My whole works depend on these cells and i cant manage to find a good stock...Please hepl...Thanks!!
MCF7 cells generally requires high glucose media.Check whether you are using it, else you have to supplement it in the media.
I work on MCF7 cells and they are fine even easy to maintain. To solve this problem try to maintain your cells in a DMEM medium supplemented with 1%P/S and 10% FBS.
How many times did you passage your MCF7 cells?
Below are two references to succesful use of MCF-7, maybe this can help.
Vanparys C, Maras M, Lenjou M, Robbens J, Van Bockstaele D, Blust R & De Coen W (2006) Flow cytometric cell cycle analysis allows for rapid screening of estrogenicity in MCF-7 breast cancer cells. Toxicology in Vitro 20: 1238-1248.
Vanparys C, Depiereux S, Nadzialek S, Robbens J, Blust R, Kestemont P & De Coen W (2010) Performance of the flow cytometric E-screen assay in screening estrogenicity of pure compounds and environmental samples. Science of the Total Environment 408: 4451-4460.
I have also made good experience with RPMI 1640 Medium. Supplied with 10% FCS, 2mM Glutamin, 1% S/P.
Our MCF7's grow well in DMEM (low glucose, 1 g/L), +2 mM L-Glutamine, and 10% FBS. 5% CO2 incubator. Are other cultures in the same incubator growing as they should? Also, MCF7 should not be split too lightly. Between passages is usually 3-4 days. If addressing these things doesn't improve your culture, I'd try to get a new vial from a trusted lab or ATCC. Good luck!
You should also check for mycoplasma contamination, which may slow the growth of MCF-7 cells.......As suggested already, you should procure a new vial of MCF-7 cell line from ATCC. Best wishes & good luck!
Check for any type of contamination, although your symptoms may well be explained by a strong mycoplasm infection on medium with rather low levels of amino acids. I've seen this problem before with HCT116 on DMEM, on McCoy's they survived.
Hi! I've experienced MCF-7 to detach, grow slowly and change morphology when I used 0.25% trypsin when passaging. Also, they have to be kept quite dense in culture and do not passage them more often than every 3-4 days. The media we use is DMEM suppl. with P/S and 10%FBS and cells look happy and grow well. Good luck!
Make sure you split your cells when they are not more than 70% confluent- I have also found that some batches grow poorly but once you establish your stock freeze down lots of batches and if this problem arises again go back to an earlier stock. I would also suggest potential mycoplasma contamination.
Thank you very much for all the answers..i grow my cells in MEM with 10% FBS and 1% PSA...the passage number is 57.
I will check for the mycoplasma contamination and will revive and adapt my cells in high glucose medium..hope that would help...thanks once again for all your answers
Please check the mycoplasma infection. There are banks which come mycoplasma contaminated . Sometimes problems is not visible on the beginning but after multiplication of mycoplasma
Hi, You can get MCF-7 cells from NCCS, Pune through proper channel. Kindly visit the website for procuring cells. I also suggest you to do passaging twice-thrice a week. If the problem exists still, try increasing the concentration of serum or the revived vial may be infected. The medium we used is DMEM, High glucose with 10% FBS. Also, your passage number seems quite average. Try with the early passage cells. Good Luck
- Badges
- Science method