I’m culturing lymphocytes for MTT assay.Can someone please tell me that once I am done with the isolation stage of the lymphocytes I.e the lymphocytes have been separated from whole blood and suspendEd in RPMI media, how long should the cell be kept in CO2 incubator before I can use them for the assay? I mean can I use freshly isolated cells, or do I need to make them grow, if yes then to what stage? So that I know that now they are suitable for MTT assay.
You may leave them in RPMI for few hours in co² incubator to make them stress free after that you can use them for your assay.
Hello Mehar Khalil
For invitro cell-based assays either on fresh or frozen PBMCs, you need to have a resting period (typically overnight) after Ficoll isolation from whole blood or cell thawing.
Best.
Malcolm Nobre Hi, what do u mean by resting period? you mean I need to put them in a CO2 incubator overnight after isolation
Yes. Leave them in RPMI in CO2 incubator overnight as mentioned by Abhishek Kumar
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