IUBio

Replacement of "old" 3H-thymidine assay.

Schrum schrum at mail.med.upenn.edu
Wed Mar 24 16:32:20 EST 1999


If you have access to a flow cytometer, you can use CFSE (Molecular
Probes).  It is a green dye that can uniformly label lymphocyte plasma
membranes, and it dilutes in half each time a cell divides.  Nondividing
cells remain labeled virtually indefinitely in cell culture, and CFSE
labeling has also been used to track cells and examine their
proliferative history in vivo.  Plus, it's a cinch to use.  See Wells,
et al., Journal of Clinical Investigation 100: 3173-3183 (Dec 1999) for
description of labeling procedure.

good luck,

--Adam Schrum
laboratory of L.A. Turka
Univesity of Pennsylvania


Jens Rainer Hansen wrote:
> 
> I want to replace my 3H-thymidine assay on human lymphocytes, since it is
> to hard to perform when having a lot of samples.
> 
> I have tryed alamarblue but it doesnt work with lymphocytes.
> The MTT assay should also be insensitive with these cells.
> 
> I have also tryed Brdu incorporation it seems ok, some times, but since the
> lymphocytes are non adherent it is nessesary to dry the cells on the
> microtiterplates with an hairdryer, and this procedure was not stable for
> me.
> 
> Do you have experience with an easy assay to meassure DNA synthesis/cell
> proliferation on human lymphocytes (non adherent) using other methods than
> 3H-thymidine, or methods where it is easy to handle the samples using
> 3H-thyminine, that means not punching filters into vials etc.
> 
> Yours Jens R. hansen



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