proliferation assays

bkleine at ruf.uni-freiburg.de bkleine at ruf.uni-freiburg.de
Tue Sep 17 08:24:56 EST 1996


>It's a lot more painful.  I would stick with tritium.
>
>Andrey Shaw
>
Whether to use 3H-thymidine (TdR) or bromdesoxyuridin (BrUdR) should depend on several points

a) safety:

TdR is radioactive and needs (in Germany and France as I know) strict regulations and a working infrastructure to use, measure and to dispose off contaminated trash. TdR is also cumbersome to measure and if you happen to have an accident at your bench, the succes of decontamination is not readily to determine: Using slabs you try to get radioactivity adsorbed, add scintillator and eventually will be able to say that you have decontaminated your bench. The half life of 3H is 12 years, to long to wait to have the place decontaminated naturally.

BrUdR is (at least potentially) carcinogenic. You do not need as strict regulations as applied to TdR, but safety regulations do apply as well.

b) infrastructure

TdR needs a very expensive infrastructure. You will need safety rooms with limited access, the licence from your regulatory body, regular health examinations of the personal. Furthermore to measure 3H you may need at least to different beta counters: One you use with your microplates and/or glass fibre filters onto which you harvest the cell cultures, another one to monitor contamination rspp decontamination. These machines are not at all inexpensive.

To use BrUdR you will need an ELISA plate washer (hopefully) and an ELISA reader. These are usually present in immunological laboratories. 

c) price

For the TdR assay you will have to buy 3H-TdR which, on a per culture base, will be less expensive than the BrUdR assay. You should also add the cost for the disposal of the radioactive waste which has tended to increase steadily in the last years.

d) conclusion

In an environnement where radioactive materials and 3H-TdR have been used, it is reasonable to continue using the TdR uptake assay for the measurement of cellular transformation and proliferation.

In a place where no infrastructure exist to use 3H-TdR, one should directly start using BrUdR, avoiding delays which would necessary to establish all the regulations and avoiding, too, the use of radioactivity.

Bernhard Kleine
Institute for Immunbiology
Freiburg, Germany
http://www.uni-freiburg.de/immuncell/bkleine




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