labelling dog muscle cells (primary culture)

David Steffen steffen at mbcr.bcm.tmc.edu
Wed Dec 23 13:49:15 EST 1992


In article <168C69C29.BIOSEE at UKCC.UKY.EDU> BIOSEE at UKCC.UKY.EDU (Stephanie Edelmann) writes:
>As an infrequent reader of this group, I don't know if this question is
>appropriate in this context.

100% appropriate, IMHO (though see below*).

>Here's my problem.  I am supposed to label a primary dog muscle cell
>culture so that the cells can be recognized when they are reinjected
>in the same dog.
> 
>1. Is it feasable (sp?) to use a marker-plasmid to do that or
>are the cells going to be senescent by the time I have transfected
>and selected them?  (I have never worked with primary cell culture)

I have not heard of this being done successfully, but I *AM*
"speaking" off the cuff.

>2. Should I go to a retroviral vector?  They are supposed to be pretty
>selective in their host.  What vectors could be used in a dog?

This is what most people do.  I'm pretty sure that the standard
amphotropic vectors will work fine in dog cells.  Muscle cells might
present problems, however, as retroviruses can only infect replicating
cells.  Myoblasts might be ok, if that will do.

>3. Where could I find information of this kind?  Not only the above
>questions, but things like "general" transfection procedures etc.

We have looked for a good cookbook on these kinds of techniques and
have not found one.  This may be because the techniques change to
rapidly.  All I can suggest is a full blown Medline search.

Once you are searching, look for the new approaches which use
asialoglycoproteins and adenovirus to increase transfection
efficiency.  This might work well for muscle cells.

* You ask for email responses due to a lack of good access to bionet.
I sympathize, but strongly encourage you to try to improve this
situation.  In the first place, this topic is one for which an
_exchange_ on the newsgroup might be better than a simple collection
of replies.  Further, once you have collected email responses, it is a
nice thing to do to post a summary to the group for the help of
others.  Finally, you would be suprised at the number of good ideas
that come from following the group regularly.


-- 
David Steffen
Department of Cell Biology, Baylor College of Medicine, Houston TX 77030
Telephone = (713) 798-6655, FAX = (713) 790-0545
Internet = steffen at bcm.tmc.edu



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