MDA-MB-231 cell culture - (Mar/01/2005 )
I've just taken out some MDA-MB-231 from liquid nitrogen. They're growing in DMEM +10%FBS + glut + antibiotics, 37'C, 5% CO2. They're not looking like epithelial cells at all. The media went cloudy, and the few cells that stuck down are really quite pathetic.
I'm growing up several cells at the moment, and they're all fine. Just these ones, and I need them desperately.
What do I do?
Help me please.
My MDA-MB-231 cells were cultured in the same condictions as u did. And everytime I thawed it, it is always fine than MCF-7 cells. As you mentioned that the media went cloudy, is there no contamination happened or something wrong in FBS inactivaction or FBS quality from manufacture as we happened before?
Hope your cells find the right way to grow!
I don't know, i'm growing other cells in the same media, and they're not acting funky at all.
Might be the cells.
It sounds like your MDA-MB-231 cell line is infected with bacteria. Do you use any antibiotics during culture? If so, how old are they, and have you looked at a drop of the medium under high magnification (400+) to see if there is any sign of bacteria?
If you have an infection ,it is possible to clean the cell line, but quite difficult and can be pretty time consuming. It is always better to get a fresh batch of cells from somewhere and start anew.
HAHA, apparently there is a bacterial infection in my cells
last year, before i was working here, the cells became infected, and a very lovely research assistant froze them down, bacteria and all. without telling me all batches are infected
I'm using pen and strep, in my media....
how can I clean them up?
You can clean them up by adding high doses of pen/step but this is always difficult-a background infection nearly always remains and as soon as you leave the media a little logner on the cells the infection flares up again. Its always best in these circumatances to thaw up a new one-maybe someone else at your institution might be able to lend you a clean vial?
I just found this page looking for info on MDA-MB-231...so sorry if I'm a little late in a reply.
I've used 1-5ul of i.v. Cipro on a plate (10cm, 10ml media) that I've needed to clean up. I only use it once or twice on the cells, and usually it clears up the contamination (Cipro is a wonder drug for sure! Yes, Yes, I use it sparingly as to not make any resistant bacteria in the lab!)
I also use Cipro to clean up mycoplasma infections but truthfully I would probably just try and get a vial from someone in another lab or just get a new one from ATCC if possible. I wouldn't want to keep infected cells in my incubator unless I absolutely had to.