Hi,
I am culturing HaCaT cells, stimulating, collecting and measuring antimicrobial peptides produced. Suddenly one time the signal was relatively low (the response according to the peptide production - e.g. 4000 instead of 30000). Then the next 2 passages gave no response. Next passage gave low response. Then I took new cells but also kept the old flask. The new cells responded only once and then the response was either low or zero. I did DAPI staining and there was no sign of Mycoplasma. After a few passages they start to grow slower.
I have been trying to find the cause of the reduced response for months, changing media, getting new cells from liquid nitrogen (from passages one year ago, when everything was fine), getting new stimuli, new Elisa kits. I still cannot figure out what is wrong. Please any ideas would be really valuable!! Thank you!!!
Best wishes,
Virginia
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