Dear colleagues,
I have a strange and persistent vacuolarization of Caco-2 cells (see the picture below), happenning after 3-4 passages with each thawed ampule. The culture medium is DMEM + glutamine, 10% FBS, NEAA and Pen/Strep.
We changed and filtered all the reagents, but the problem persists with cells from various sources.
We could not detect any bacterial infections by microscope or laboratory tests, and all viral PCR-tests were also negative.
Do you have any ideas of unusual bacteria, funghi or virae to be tested for?
Have you ever experienced such reaction to any reagent or medium supplement?
For subculture, we use 0,25% Trypsin-EDTA. Could the vacuolarization appear due to regular trypsin treatment? Is there anything we can do in this case?
Thanks a lot.
Hi Nadia,
They are also known as 'domes.' I believe they are normal in a cell line such as Caco-2. They are also found in other epithelial cell lines, especially those that develop epithelial barriers, such as MDCK cells in culture. If I recall, they are the result of cells forming junctions and a barrier (either 100% confluent or clumps of cells that have formed a monolayer). If anything, they are a positive sign that your cells are functioning similar to gut epithelial cells would with regards to barrier formation.
Also, if you are studying their ion channels, or barrier, they should be grown and analyzed on transwells. The number and size of vacuoles will likely be reduced.
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