I have problems with adhesion of human epidermal keratinocytes (HEK) cells to the bottle. I defrosted one ampoule of this cell line a week ago in a T-25 ready for adherent cells, then I had to expand to a T-75 flask. So far, the cells grow normally, but I don't see any adhesion nor change in morphology.
Do you have any advice?
IS there any sign of contamination??? did you checked prior that the cells you thawed are healthy enough when freeze?? Also now you can try one thing, just divide your culture in two parts and seed one in T25 and other in T75 in high confluency see whether you can have a solution or not??
Hi Anabel,
when you say you had to "expand" I assume you mean that trypsinised the monolayer.
If so, perhaps you overtrypsinised and/or did not fully inhibit the trypsin, affecting adhesion.
Adherence is always a problem for keratinocytes. For better results you need to use coated culture flask not the normal TC treated ones. I found CellBind surface from corning is good for keratinocytes. BD biosciences also provides specially coated culture flasks, for keratinocytes the carboxyl coated cultureware is best. As I also worked on keratinocytes and I am fully agree with your observation that in normal TC treated cultureware the keratinocytes didnt adhere efficiently to the surface. Just use one of the above mentioned coated flask and your problem will be solved. Good Luck
- Badges
- Science topic