In article <01H0S46AE9UA0077YU at GW.AGR.CA> RAMPITSCH at BCRSSU.AGR.CA writes:
>>Hello nets:
>I'm not sure if this is the right place to ask, but .....
>I have a hybridoma cell line contaminated with fibroblasts.
[stuff deleted]
>Now I have a line of fibroblasts which refuses to die...
[more stuff deleted]
>Is there a miracle anti-fibro-
>blast agent? Is there any way of selectively getting rid of these fibroblasts?
>>Thanks for any help and comments,
>Chris R.
>
Chris,
I'm not an immunologist, so I welcome any comments or criticisms on this
attempt at an answer. The two methods I have seen for killing fibroblasts
are:
1) culture medium with d-valine substituted for l-; fibroblasts lack the
enzyme to utilize the d- isomer. Available from Gibco.
2) anti-Thy1.1 followed by complement-mediated cell lysis. Don't know if
this would work in a hybridoma culture. Thy1.1 is a fibroblast-specific
cell surface marker. I can look up the antibody source if you want.
Am I the only person on earth who has killed fibroblasts? I forgot to add
pen/strep to a batch of DMEM/serum, but I bet that isn't the answer you're
looking for :-).
Reference to the above procedures (except forgetting pen/strep): Levison
and McCarthy, _Culturing Nerve Cells_ (Banker G and Goslin K, eds),
MIT Press, 1991 (I think).
Hope this helps
Jim
--
Jim Hutchins [] E-Mail: jbh at anat.umsmed.edu
Asst Prof of Anatomy [] Asst Prof of Neurology
Univ Mississippi Med Ctr [] Jackson, MS
