I´m culturing Hela cells. In the flask and normal petri dishes they look wonderful, but when I seed them in glass bottom dishes they just die. I tried different dishes and I always get the same problem. Any suggestion?
We also have a feeling that they attach badly to chamber slides (especially after transfections). You can coat with collagen or poly lysine as Dr. Claro mentioned above or even keeping some high serum medium for few hours on dishes before adding cells also could help.
Poly-L-lysine is certainly very useful. If using coverslips, I get the impression that cells like some brands more than others. I got good results especially with Assistent/Hecht. It may help in all cases to pretreat the coverslips with Ethanol/HCl, then wash carefully with 70% Ethanol to remove the acid.
If using glass bottomed dishes- they can be quite dirty. I used to place them in a glass jar, filled with ddH20, with a few drops of versa clean (Fischer), and sonicate them for 20 min. Followed by several rinses in ddH2O, until the detergent bubbles are gone, then sonicate in ddH2O for a further 20 min. Rinse 3 times in ethanol and air dry in the TC hood.
my cells also required fibronectin coated dishes.
Thank you all. Very nice advices.
But I think I don´t have problems with attachment, just with survival.
I´ll try the cleaning methods.
Thanks
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