I expose lung cancer cells to hypoxia to study gene expression. The cells were kept in a hypoxic environment for 6 hours, and after 6 hours, I lifted the cells using trypsin and left them in the incubator for 3 minutes. In this case, have the cells transitioned from hypoxia to normoxia? I would appreciate any guidance on what I should do in this situation
Good question! Trypsin treatment does not lead to cell death, so living cells are in contact with the trypsin solution for a while before being deoxygenated again in the incubator, time enough for undue cell reoxygenation that may trigger lots of disturbing reactions. I suggest deoxygenating the trypsin solution before bringing it into contact with the cells, for example by placing the trypsin flask (opened!) in the same incubator for a while before use.
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