how to prevent HEK293 cells from coming off the plate - (Mar/08/2005 )

I am working with HEK293 stably transfected cells, the problem right now, they are easily come off the plate. I already tried using collagen coated and poly-L-Lysine hydrobromide coated plate, they are still detached easily. When I use cells for binding study at 4oC, and during washing, almost all the cells detach from the plate.

Can you give me any suggestion for treating HEK293 cells?

Dairil

hi
maybe you can try to change the plastic type of your plates. i know there are differents types that may increase the attach. But i was wondering, do your cells still dividing themselves?

Hi,

During incubation at 4oC, I guess they are not dividing anymore. However, when I plated them until I used them for experiment, they are still dividing.

Do you know what kind of plastic type of the plate that I can use, right now I am using plate made by polystyrene.

hi
i good quite bad news for you. in my lab we use polystyrene too. So the problem may not come from the plastic.
But the bad news is that my colleague just told me that she had worked with 293 cells, actually different from the one i work now with, that were very few adherent on the plate. she says that it depends on the lineage of the 293....

Maybe you can try others 293 in your area.
Sorry for the news.
I wish you good luck.

Fred

I used fibronectin-coated plates for growing 293 cells, and it worked quite well. When plating them on nude plastic half of them dettached easily, but not from the fibronectin.