I have to differentiate 3T3-L1 and further investigate the cells with MTT assay and scratch assay. I need to confirm something regarding the cell after undergo differentiation. Can they maintain as adipocyte cell after trypsinized and sub-culturing or the differentiation should be done in well plate to proceed with MTT assay and scratch assay. Thank you.
Once the 3T3-L1 preadipocytes were differentiated into mature adipocytes (in-vitro condition) they can be used for final experiments (eg. MTT assay). These adipocytes can not be subcultured. I think scratch assay will not be the right experimental choice with 3T3-L1 adipocytes, as they are terminally differentiated cells and will not divide. Eventually, we find trypsinized adipocytes lost their ability to attach to the matrix when it was seeded with fresh culture media.
Thank you so much Devarun Patra for the explanation. It's meant a lot for my study now.
You can directly use your mature adipocytes to check cells viability by MTT assay. There is no need to trypsinize the mature adipocytes for subculturing. For subculturing, you need preadipocytes in 50-70% confluency while mature adipocytes don't proliferate.
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