[Cell-biology] Re: Lung fibroblast methotrexate toxicity

PiLS via cellbiol%40net.bio.net (by pils from invalid.ca)
Thu Jul 12 15:53:41 EST 2012

Sonal Patel <stylosweetu from gmail.com> à ploppé:
> I am trying to find out effect of methotrexate on lung fibroblast WT cells
> and KO where gene of my interest is knocked out . After 3 weeks on staining
> with crystal violet I get lot of background cells in KO alongwith colonies.
> Can anyone suggest me how to remove the background cells.


can you provide more info about what you are trying to achieve? Are you
using the MTX as a selection agent or are you trying to study its 
effects on KO cells? 

In general if you have incomplete selection (assuming that's your
problem) reducing the concentration of serum and/or increasing the
concentration of the selection agent gives good results.


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