I wonder if anyone has cultured THP-1 cells in shaker-flasks?
If so, what is the appropriate shaking rate? And if you recommend any anti-foaming reagent being added to RPMI? Or, do you recommend another kind of medium?
I also wonder if incubation in an 8% incubator (vs. 5%) might cause any problem.
It should be possible to culture THP-1 cells that are in suspension on a shaker. I would not change the medium, control the speed (keep as low as possible), and keep CO2 levels to 5%. However, due to alterations in culture conditions, cell signaling pathways that determine cell survival are likely to change. Therefore, unless there is a compelling reason to change the culture conditions, as a control, one may have toi culture cells as recommended and compare with the new culture conditions with respect to alterations in cell signaling pathways.
Thanks Divaker;
Is there any specific pathway that you would recommend to check for this? i would appreciate to know exactly which protein and test you recommend.
I would check for NF-kB activation and AP-1 activation. Both regulate a number of genes. Therefore, you have lot of choices to identify genes for which you have qPCR or PCR assays.
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