I am performing an assay on my adherent cells and up until the day of the experiment, the cells grow nicely and reach a good confluency of ~ 80 % cell density. However, on the day of the experiment, after washing in PBS and gently adding the media, the cells start to dissociate. The media and PBS are both pre-warmed to 37 C. Could this be associated to the cells being overconfluent?
Hello
What is the percent FBS you are using in the culture medium?
You have not mentioned the culture medium you are using.
Best Wishes.
Dear Saeed!
Of course, your cells will dissociate at 80% confluence. Moreover, cardiomyocytes are a very demanding culture. Cells are weakly adherent and strong adhesion flanks are needed to grow cardiomyocytes.
Hi, did you coat with gelatin your plates or flask? As suggested above it can be an issue. I worked with HL1 and coated the plates
Valeria
Malcolm Nobre Malcom, for the A16 cells, I use 12.5 % FBS to grow and expand them. Valeria De Arcangelis , I did not coat the cells, as they seemed fine for expansion, but perhaps for this particular assay, using Gelatin could be a good suggestion, thank you! As for flask, our lab generally uses cell culture dishes, but I have used flasks in previous flasks, but would using flask make that much a difference? Thank you all for your suggestions!
Hi, flasks are only to expand clone or if you need to process a lot of cells in one experiment. If the cells grow well use plates and if you choose to use gelatin the plates are much easier to coat
Valeria
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