293 Cells: how to transfect?

jstrassw at opal.tufts.edu jstrassw at opal.tufts.edu
Tue Jun 25 10:23:21 EST 1996


Hello!
Thanks to all who gave me recommendations about 293 cells.  Some replies were
by email, I'll tell you what they said:
1) Dont wash with PBS/EDTA (as we usually do post CaPhos transfection.  They
need the divalent ions or will detatch immediately.
2) Use FALCON plastic (we do)
3)keep everything at 37, not room temp.

SO, I did thei and my problem is solved!
Thank you all!
John




> HEK293 cells are notorius for not sticking to anything! We use them to 
> transfect with Glutamate receptor subunits and stick them to glass 
> coverslips coated with poly-D-lysine. They seem to be OK then. They can 
> be pathched, used in confocal microscopy and immunocytochemistry (live) - 
> so they will stick! But they are very sensitive to their environment. 
> They don't like being at room temperature when growing on ordinary tissue 
> culture plasticware and can die very easily if they don't like your 
> transfection methodology. We use a CaPO4 method, with overnight 
> transfection incubations, and they seem fine with this.
> 
> Hope this is helpful
> 
> Andy Doherty
> 
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