2 replies to this topic

[biolcrazy]

Hi,

I have been culturing MDA-MB-231 cells for the past couple of months on DMEM+10% FBS. I got these cells from another lab but I have no idea about the passage number. When I got the cells from them I was told not to worry about the passage number as these are cancer cells and they will continuously keep growing. Initially when I was culturing these cells they were fine and I had no issues.

However for the last few passages I have observed that I am getting an increasing number of senescent cells i.e. first there were one or two per plate but as I am passaging them they seem to increase in number-- see attached pics.Has anyone had a similar problem? Should I just go ahead and buy a new batch of cells???

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[bob1]

How long have you had these cells actively growing?  The longer you have them up, the more they change from the parent strain and, despite them being "immortal", the more likely they are to start undergoing senescence.  You should thaw a fresh vial.

[Curtis]

I have had problems like that bofore. Eventually those cells die and most of the time I had senescent-free flasks after a while. Their increase is not only because of shortened telomerase, but also due to the media you use. DMEM is my favorite medium, but ATCC recommends Leibovitz's L-15 Medium for your cells.We also use less than 10% FBS. what is your antibiotic?