Hi,

I am growing Caco-2 cells on PDL coated coverslips for immunocytochemistry. Most literature mention the cells require 21 days to differentiate fully. However, I’ve noticed that my Caco-2 cells become confluent within 3 days of seeding.

Does anyone have advice on how to maintain these cells for the full 21-day differentiation period without overgrowth or compromising their differentiation? Should I adjust seeding density or explore other strategies?

I tried 5* 10^4 cells/ ml, 1* 10^5 cells/ml and 3*10^5 cells/ml.

if you have a working protocol for growing and differentiating Caco-2 cells under similar conditions, I would greatly appreciate it if you could share it.

Thank you