I am testing the intestinal permeability of some compounds using Caco-2 monolayers on Costar 3460 Transwell Permeable Supports. I have been told that you might re-use the supports, taking the cells out and starting again. Has anyone done this before? Does trypsin damages the support? How many times can you re-use the support?
Hi Laura, I have not done permeability assays using transwells, but sometimes I reused transwell inserts for cell migration assays.
I used Q-tips to gently ablate the cells from the top and bottom of the inserts, put them in 1 M HCl overnight at 4oC and then washed them with sterile H2O and dried them.
It is not advisable to re-use it. I tried it once, but the cells did not adhere completely on the membrane, and there were some gaps, especially at the boundaries of the wells. It may work for you, but if you want to save time, take a new one.
If you just want to exercise/practice and set your hand for the manipulation, then you can re-use it. But if it is a serious manipulation, dont take risk, use a new insert.
All the best Laura
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