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Dear all,
I am facing some problems with Caco-2 cells. I am using the classical
21 days protocol and everything was working well, recently, however
after 18 days of culture over transwell membranes the cells start to
detach. I used the passages over 56. I thought it was probably
because the cells were too old so I purchaged new cells, seeded in
33-38 passage. They also detach at 18 days. I usually determine
resistance after seeding two weeks, the resistances are over 400 in
14 to 17 days. At 18 days, they start to detach. I do change medium
in 2-3 days. I cannot see contamination, just feel cell are not
strong, fade away. I tried cell density from 70,000 to 100,000, the
results were the same. Did anyone have similar problem and may help me?
Sincerely,
Whocely Victor de Castro
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The following message was posted to: PharmPK
I had similar problem when I used one type of 6-well plate to grow
Caco-2 cells, but the same cells grew well in 100 mm dishes. So I
think your problem may be the transwell, not the cell itself. You may
try a new ordered transwell with the previous one to compare.
Hopefully, this can solve it.
Shanjun
University of Toronto
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Dear Castro
The problem might be with the transwell plates.
I had a similar problem with a lot of TC flasks the cells were
getting detached after a confluency of around 60-70%. Please try
changing the transwell plates
Hope this helps
Naveed
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