help: MB-231 - help (Jun/02/2005 )
Could anybody tell me how much generation is maximum for MB-231 cell line research. Mine is 50 generation already, and cannot repeat myself very well by using this P50 MB-231 cell line. I want to know if this cell is too old for research, do I need order new cell now? Thank you for everybody's help.
i thought that with most cell lines you should stop at ~20.
Hi,
They are immortalized. Passage number should not matter. In fact, if you order from ATCC, the passage number will be over 100.
Passage number does matter if you are culturing primary. e.g. HUVEC cells shall not go over p5.
Good luck.
They are immortalized. Passage number should not matter. In fact, if you order from ATCC, the passage number will be over 100.
Passage number does matter if you are culturing primary. e.g. HUVEC cells shall not go over p5.
Good luck.
Depends on the cell line. Sometimes characteristics are lost when the cells have been passaged too many times. This is not the case in all lines. The best idea is to always have a vial frozen in stock, so IF you've lost certain characeristics, you can whip 'em out.
Of course it depends upon the cell line. That is why I mentioned HUVEC (I used them for cell-cell adhesion experiments with breast cancer cells). But, MDA-MB-231 is very strong cell line along with Hs578T and DU4475 (all ER negative cell line). I kept those cell lines in -80 deg. C for over 2 years, but as far as I've seen, they have no problem sub-culturing. Although ER negative, MDA-MB-468 did not like in -80 C for long time. Most of the cell were dead when I tried to sub-culture them (so as MCF-7, SK-BR, ZR-75-1). T-47D, which is ER positive cell line, didn't mind -80 C.
I'm at home, so I can't tell you exact passage number of MDA-MB-231 when I ordered them 2 years ago, but I think it was passage 134. If I have time tomorrow and can find a space to upload an image some place, I can scan and post MTA of MDA-MB-231 when I received the cells.
I think passage number is only so you can go back and repeat your experiment at the same point in time. I have cell lines that are in the upper 290s in passage. As long as you do the same experiments in the same range of passage, it is fine. Cell lines definitely change over time. So comparing experiments you did at pass 4 with experiments you did at pass 44 is not going to be all that valid, in my opinion.
Beverly
Lipo2000 as protocol from invitrogen works well for mcf7...though incubation time needs to be extended to 10hrs instead of 4-6hours. 231...not sure.