Recently I'm facing a problem in my cell culture. Lymphocyte-like cells (K562 cell line - suspension) are adhering to the flask and thus I can't get the cell concentrate after centrifugation.
Should I trypsinize these cells?
Is it possible theses cells changed their genotype due to the number of passages?
I await a help,
Thank you!
Hey Caio,
three possiblities:
1. depending on the amount of passages they might have mutated. I always rethawed my cells after 15 passages.
2. Your lymphocytes might have been activated by a subtle contamination and therefore they start to adhere.
3. Did you change your serum (the one for the cells not actually yours :) ) recently? This can contain activating substances.
3. Are you using medium, trypsin and wash buffer just for those cells or also for other cell lines? Might be a contamination with another adherent cell line. This is the worst case though
Good luck,
Teresa
Hey,
K562 cells can spontanously differentiate. If you did not treat them well (eg not passsaged often enough, medium wasn't change often enough, wrong medium, too less serum...), more differentiation can occur and then it is possible, that they attach. Do NOT detach them and continue with the culture. Thaw and culture a new vial of cells with a much lower passage number.
And I agree to Teresa, it is also possible that they can mutate, they have a quite instable genome. Another reason to go for a fresh culture with lower passage.
Good luck,
Floriane
Thank you so much for the answers.
I'll try to thaw a new vial and use new and fresh media, serum and trypsin to get a healthy culture.
Best regards!!
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