G418 and resistant cells - can't kill the cells (May/18/2007 )
Hello everyone,
I'm trying to establish stable cell lines with different cell types. Some of them seem to be naturally resistant to g418, ie: I can't kill them even with very high concentration (from 0.1 mg/ml to 5mg/ml) of g418. I did a kill curve to check that, I even tried to dilute them more and see the effect of g418 on more diluted cells. Still I didn't succeed to kill them. And the g418 works apparently well because I can kill N2a cells in less than 10 days.
Is that possible that the cell type is naturally resistant to g418 or could it be because of something else (growth rate...)?
I need your help
thank's.
I'm trying to establish stable cell lines with different cell types. Some of them seem to be naturally resistant to g418, ie: I can't kill them even with very high concentration (from 0.1 mg/ml to 5mg/ml) of g418. I did a kill curve to check that, I even tried to dilute them more and see the effect of g418 on more diluted cells. Still I didn't succeed to kill them. And the g418 works apparently well because I can kill N2a cells in less than 10 days.
Is that possible that the cell type is naturally resistant to g418 or could it be because of something else (growth rate...)?
I need your help
thank's.
some cells are resistant to G418, not naturally though. Make sure about the cell lines before you proceed.
I believe some neural cells are naturally resistant if I remember correctly. Yes, the speed of killing depends on the growth rate.
the ability of cells to act against toxins is different, you have MDR genes such as P-glycoprotein, cytochrome P450 etc which are differently active in various cell types...
Yes some cells are resistant to G418.
how long did you culture the cells in G418 for before deciding they were resistant???
you need to dilute well your cells as they can divided well. G418 acts only on active (so mean in general dividing) cells
Thank you all for your answer. Looks like I won't be able to use these cells in my experiment. too bad.
To respond to lauralee, I cultured my cells for 3 weeks under g418 pressure, and still was'nt able to kill them.
Bummer!! they must be hardy little critters 
Goodluck with finding another way to get the answers you need