Basically i need to remove some monocyte macrophages from a 96 well culture plate to store them for later use, which may well be cellular gene expression analysis or more simple viral DNA quantification using PCR. Any help is most appreciated, particularly with regard to the removal of cells from tiny wells. If anybody has protocols to share :) thanks
Macs are sticky cells. We have used the following to dislodge our mature APC from polystyrene:
Ice-cold DPBS no cations to cool cells and cause retraction of integrins
1x TrypLE from Thermo-Fisher at 37*C for 10 minutes to break peptide adherence bonds without damaging the membrane
pipette, spin, decant TrypLE, resuspend in warm PBS until ready for RNA later.
In a 96-well plate you could try just the TrypLE, but spin and remove right away our viability will take a hit
Cheers!
You may wash your adherent cells with cold DPBS followed by adding RNAlater to well. Pipette it well (multiple times) to dislodged the cells. Look into microscope for non adherent cells and store the suspended cell in RNAlater.
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