We are currently using the air-liquid interface technique to differentiate human primary nasal epithelial cells. While the cells are differentiating well and producing cilia, some are also generating a lot of mucus. We need to remove the mucus for downstream techniques such as immunofluorescence staining. We have tried washing the cultures with PBS for 10 minutes at 37°C, repeated three times, but this only partially removes the mucus. We fix the cells with 4% PFA.
We would appreciate suggestions for other solvents that could be effective in removing the mucus layer. Please note that we have already tried washing with PBS and Tween20, and it only partially removes the mucus. Any insights would be greatly appreciated.
Hi Aquib,
for washing, Try using reducing agents like DTT in combination with low amounts of 0.01-0.1% CHAPS detergent (maintain osmolality with NaCl)
post to wash remove wash buffer with PBS
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