Recently starting working with HEK cells. I noted that after my medium change (10Cm plates, suction of old media, and adding 10mL of new complete media) my cells all started to float. I now that HEK cells don't stick well but I wasn't expecting such a drastic detachment with simply changing of media. Solution/suggestions?
What you describe is the normal behaviour ob HEK cells... I also suggest to coat the plates with Poly-Lysine. Since we started doing this we didn't have any problem anymore...
When I use HEK293 cells I coat the plate with Poly-L-Lysine.
1 well of a 6 well plate would require 0.5ml of poly-l-lysine at 0.1mg/ml. Remove and wash the wells in PBS then leave for a few hours to dry.
I would also use the poly-l-lysine several times before discarding it.
I could then use 500,000 cells/well of a six well plate for several days without loss of cells.
What you describe is the normal behaviour ob HEK cells... I also suggest to coat the plates with Poly-Lysine. Since we started doing this we didn't have any problem anymore...
I too have seen this monolayer behavior with Hek293 cells.
They attach but very loosly. If the culture flask gets bumped i have seen the whole culture come loose and float up.
When i was doing medium changes i would have to very carefuly replace medium by pipetting slowly along the flask wall.
Best wishes
- Badges
- Science topic
- Similar topics
- Physical Sciences
- Materials Science
- Plating