I need the cells to adhere to the plate for a fluorescence assay. Growing and sterility aren't problems. For some reason, once I move the reconstituted cells to the well plate, they stop sticking.
My media is RPMI with 10% FBS, 1% PennStrep, and 1% Glutamine
(It's not the best but they're proliferating like crazy in the T-25 culture flasks so I haven't had any issues here).
The well plate was UV treated for minimum 3 hours but was was for general fluorescence assays.
Dear Frederic,
Three possible explanations:
- the plate is not tissue culture coated.
- coating does not allow hct116 cels to adhere (unlikely).
- trypsinised cells are dead.
You need to seed an aliquot of the trypsinised cells in a plate/flask where you know hct116 cells grow well and to use in parallel a second cell line as a control. This test will be useful to clarify the unexpected behavior of your cell line.
Good luck!
Metello
You can also try to pre-coat the plate with poly-lysine or collagen which will facilitate the attachment.
@Eugen could you share a protocol for optimizing the concentration of poly lysine or collagen?