Stable Transfection of T Cells

Frank O. Fackelmayer Frank.Fackelmayer at uni-konstanz.de
Tue Jun 8 10:32:53 EST 1999


Hi John,
I have stably transfected Jurkat T-cells recently. After testing a lot
of different transfection methods, I found that Qiagen´s SuperFect  is
the only one that works reproducibly (no affiliation). Even with this
stuff, transfection efficiencies are very low with T-cells (>=1%
transiently transfected cells), but with G418 selection it was possible
to obtain quite a number of stable clones. Transfection with liposome
based materials (Fugene, DOTAP, LipoTAXI) did not yield transiently
transfected T-cells at all (but work very well for other cells!) and,
consequently, no stable clones.
Thus, I would first try with Superfect.

Hope this helps,
Frank


Alan J Watson wrote:

> Greetings.
>
> I need to prepare some stably transfected T cells (three
> different CD4+ lines, if that matters), but I have never
> transfected this type of cell before.  Any tips on lipids or
> other reagents that work well on T cells in general, or do I have
> determine the optimal reagent for each cell line experimentally?
>
> Thanks in advance,
>
> John Watson
> Bristol-Myers Squibb Lead Discovery Molecular Biology
> watsonaj at bms.com




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