Transfection of primary fibroblasts?

Martin Leach leach at bu.edu
Fri Apr 28 09:16:28 EST 1995


>Hello netters,
>
>Does anyone have some advice for transfecting primary mouse 
fibroblasts? We
>have been using a standard calcium phosphate method that has high 
efficiency in
>the adherent cell lines that we usually use (NIH3T3, BHK). However, we 
have had
>very poor recovery of transfected substrates from our primary mouse 
fibroblasts.

Are you assaying the transfections or trying to isolate the DNA 
following transfection..?

Anyhow, using standard calcium phosphate technique I have transfected 
into a whole mess of primary cultures..including chick and rat primary 
fibroblasts. Maybe the problem with mouse is a species/tissue thing. Are 
you changing the culture media prior to transfection? Is it defined 
media? If defined..maybe you should pre-incubate the media for 24 hours 
before use (a trick that makes some cultures transfect!!! dont ask me 
why!)..

good luck

Martin


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