GFP expression of sorted cells

From: Jochen BARTHS (jbarths@titus.u-strasbg.fr)
Date: Fri Apr 19 2002 - 07:38:47 EST


Dear Flow-ers,

We recently encountered some problems with the sort of transfected cells.

When we sort GFP+ mouse embryonic stem cells (48 h after Xgen transfection
with a vector expressing the gene of interest and GFP), the sorted positive
cells gradually loose their fluorescence within 3 to 4 days.

On the other hand the same transfected cells, selected for example in G418,
retain their fluorescence much longer.
Viability etc. of the sorted cells is very good.

Any ideas, how this phenomenon can be explained, would be appreciated.
(Was there any deleterious effect of cell sorting on long term protein
expression described before?)

Any proposals how to get around it would be even better.

Thank you very much in advance.

Jochen

Dr. Jochen BARTHS
IGBMC
Service Commun de Cytometrie en Flux
1, rue Laurent Fries
BP 10142
F-67404 ILLKIRCH CEDEX
France
Tel.: +33 / (0)3 / 88 65 34 27 or 28
Fax: +33 / (0)3 / 88 65 32 01
e-mail: JBarths@IGBMC.u-strasbg.fr



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