I want to perform MTT assay for BJ-5ta and HaCat cell lines. I want to know before performing the actual experiment how many passages should be done of the cell lines?
Hello Noor Ul Ain
The cell lines used for any cell-based assay should be of the early passage preferably (between P15- P25). They should be healthy, and you should use the cells when they are in the exponential phase of growth (i.e., when the cells are actively proliferating). If the cells are already in culture and they look healthy, you may go ahead and use them for the MTT assay. But if the required cells are stored in liquid nitrogen or frozen in -80 degree C, you need to thaw these cells and subculture them for a few passages (preferably 2-3 passages) before you use them for the MTT assay. However, some cells may take beyond passage 2 to revive completely after thawing.
So, cells used for MTT, or any other cell-based assay should be in good healthy condition (morphologically and actively proliferating) to enable one to obtain reliable results.
Best.
Malcolm Nobre
Thank you so much for advice. I am on 5th passage number. Cells are growing actively. Should I perform MTT assay then?
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