Concentration Hygromycin for HEK-293 cells?

Lloyd Graham lloyd.graham at REMOVE-MEmolsci.csiro.au
Fri Nov 20 01:34:29 EST 1998


In article <3653f179.9317838 at news.cc.ruu.nl>, t.vink at med.uu.nl (Tom Vink) wrote:

> Hi,
> 
> I am stably transfecting HEK-293 cells with ahygromycin selection
> vector (pCEP4). Anybody knows what concentration hygromycin is
> suitable for theses cells ?
> 
> Greetings Tom

Dear Tom,

The published value is 200ug/ml, but I prefer 100ug/ml. With the lower
concentration (and perhaps even with the higher one) the transfected cells
grow faster than the untransfected ones die, so I harvest the cells and
re-seed them a few days into the selection. This gives the transfectants a
clear field, since untransfected ones generally don't re-adhere. 

You might like to pre-coat your dishes/flasks with bovine fibronectin, as
I generally find that selection causes these cells to become less
adhesive. Even with FN coating, a proportion of transfectant progeny cells
is non-spread and easily detached.

Hope it works for you.

Lloyd.

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