I was curious as to what technique other people use to change out media from the inner chamber of Corning transwell inserts. It can be a bit of a challenge not to disrupt the cellular monolayer when performing this task. I have used both a pipette to accomplish this and also a vacuum pump system. I am currently conducting monolayer permeability studies with HUVECS later to be used with hCMEC/D3 BBB EC cells in the near future. I am using a 6.5mm insert for a 24-well plate with a 0.4uM pore size. I was curious if anyone has figured out a reliable way to ensure monolayer integrity.
Dear Chris
Use suction method to take out the media. but be careful while sucking out the media and suction pressure should be very low. Take out the media from insert"s side wall, don;t suck media from the center of the insert. After sucking out the media immediately pour the new media from side wall, other wise cell will become dry.
Every step you should have to perform slowly other wise it will disturb mono-layer of cells.
It should be OK to suck out the medium using 1 ml pipette tip, and doing it quite slowly, from the side wall.
if you have forceps to help you, you can use it and tilt the insert one side and have your suction pump or pipette aiming at the wall to remove the liquid.
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