Hi all,
I am planning to initiate a migration study that will make intensive use of transwell-like culture inserts of various pore size. However, given the high cost that would represent buying new inserts all the time, I was considering recycling them. Do you know if that can work, without clogging the pores or damaging the membrane? And if so, what could be a good protocol for that?
thanks a lot,
Pablo
Hi Pablo: Personally, I would discourage you to pursue this, as the cost of these inserts and the "cost" of your time and the risks of contamination/inconsistency for your new data collection is too high. If you mention such a recycles use in your methods for publication, you may be questioned by the reviewers....
I've worked with transwell inserts before (with cancer cells and PBMC), and I don't think they are a very good assay system. I think the transwells only allow you to see any subtle chemoattraction or chemorepulsion.
I also don't think transwell chambers are re-usable, as you'd have to clean the small pores. You could try washing the transwell chambers in EDTA or PBS, and storing them in sodium azide, but I think that's a bad plan.
Have you considered using a more robust assay system? Insall chambers and Ibidi chemotaxis chambers allow you to record the movement of cells in two and three dimensions over broad time ranges (1-24 hours), and return much crisper data than do the transwell chambers.
The Ibidi chambers listed in the link below could probably be repurposed for a small "high-throughput" assay, though you might need several cameras.
http://ibidi.com/applications/chemotaxis/u-slide-chemotaxis-2d/
Our company develops 3D cell barrier culture systems for in vitro study of virtually any cell barrier, and the system comes equipped with both a Fluid Perfusion Unit and a Trans-Endothelial/Epithelial Electrical Resistance (TEER) Measurement Unit! The use of a vessel-shaped 3D environment has been proven more effective than 2D systems like Transwell (
Article Santaguida S, Janigro D, Hossain M, Oby E, Rapp E, Cucullo L...
), and our advanced TEER Measurement Unit allows for frequency sweeps between 0.1-1,000 Hz, automated time point sampling, logging of data to Excel, and additional measurement of phase angle for cell capacitance calculations! For those that are committed to Transwell use, we also have a TEER Measurement Unit that is compatible with nearly all Transwell products (Endohm cell cup chamber, STX2 "chopstick" electrodes, etc) and allows the user to greatly expand their testing capabilities with Transwell equipment. Additionally, we have smaller modular systems that connect to microscope-friendly cell culture modules: i.e., a miniaturized 3D cell culture system that you can use right under your microscope! If you or anybody on this thread is interested in learning more, I encourage you to visit www.flocel.com or email me at [email protected], thank you!
Hi Pablo,
Yes, you can re-use the transwells! I just finished my experiments. It works. Collect old inserts and wash them in a glass with cell culture grade detergent, on magnetic stirrer, with very gentle mixing. You may leave them for the day or overnight. Then rinse them multiple times with water and 70% ethanol and leave 2+ hrs in cell culture hood under UV light. No problem, no contamination. Good luck!
Hi Pablo, Well done Evgenia. May I ask you about the detergent you used and its concentration? Thank you.
I came across this paper while looking for an answer for myself: Article Reuse of Cell Culture Inserts for In Vitro Human Primary Air...
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