Fibroblasts won't restart after freeze

D.K. DK at no.valid.address
Wed May 7 21:59:29 EST 2003

In article <c09b237b.0305071848.6bea6062 at>, ladasky at (John Ladasky) wrote:
>Hi, folks,
>I am growing six primary human fibroblast lines.  I have vials of all
>of them in liquid nitrogen.  When I thaw these vials, four of the six
>will eventually start to grow.  The other two have viable cells but
>remain stubbornly quiescent.  I can pass them and they will re-adhere,
>but they have no interest in dividing.  They were frozen down at the
>earliest possible passage number, below passage ten in each case.  Two
>of my six lines that regrow nicely were actually frozen down above
>passage fifteen.
>Does anyone have any advice for coaxing a few cell divisions out of
>these lazy cells?  Conditioned medium, perhaps?  More FBS?  Human
>serum?  FGF?

Sounds tough. I think your best chance would be conditioned medium 
from those primary lines that do grow. 

Just out of curiosity - when they adhere, do they spread nicely 
or just sit there? 


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