I are working on HUVEC culture. 5 days ago, my HUVEC cell got confluent and my boss let me to continue grow it for a test later and I replaced the media in two days( day 3). Today( day 6),I found most of the cell already peeled off from the wall of flask, I think the overgrowth of the cell cause the peel off. Did this happen to you? Thank you!
Hello Ying Yin
Cells usually start peeling off once they are overconfluent. There are difficulties encountered while culturing HUVEC cells. They should not be allowed to become overconfluent as in your case. These cells must be passaged when they are at 50%-60% confluency.
Regards.
Hello Yin,
What you described is a typical scenario when HUVEC (and most of the primary vascular endothelial cells) are allowed to stay in culture for extended period of time. Depending on the experiment (e.g. if you need to have a good and mature intercellular junctions for assessment of the monolayer integrity/permeability either by IF or functional assay) you may allow HUVEC to grow for a couple of days post-confluence. How fast/slow HUVEC will start detaching from the dish will also depend on the passage number. Usually "younger" HUVEC (e.g. passage 1-3) can stay confluent for longer time, while "older" HUVEC tend to detach much faster once they reach confluence.
You should talk to your supervisor and clarify the experimental endpoint(s) and reasoning(s) for why you have to culture HUVEC for 6 days!?
Best of luck!
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