How to detach MDCK cells from flask while retaining cell surface cadherins
- Wash them first in PBS +4mM EDTA.
- Trypsinize MDCK in 0.05% trypsin +EDTA (2mM, standard from GIBCO), Here lose all cadherins from the membrane.
- Then let the cells recover from trypsinization in suspension (in DMEM + 10% FCS, about 10 Mln cells/ml) for 1 hr
- Wash once in DMEM + 0.5% FCS and resuspend in DMEM + 0.5% at the right concentration for use.