I am currently culturing the 3T3-L1 cells, and trying to differentiate them. What happens if I change to differentiation media immediately after confluence? Why do we have to wait 2 days post-confluent to change the media? Wont the cells be overconfluent and die off?
Hello Dayana Cheong
Yes, it is necessary to wait two days post confluence before changing to differentiation media. The cells will undergo a pre-adipose to adipose-like conversion as they progress from a rapidly dividing state to a confluent state and then to a contact inhibited state. Two days post-confluence, the cells will be balled up and start looking a little ugly, ready to be in a state to differentiate in the differentiation media containing IBMX, dexamethasone and insulin.
Best.
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