Hi everyone,
I am currently working with THP-1 cell line. I am trying to differentiate them and characterize cell population between monocyte versus macrophage. I did a basic light microscope image analysis but am looking for additional techniques. We do not have flow cytometry available. Has anyone done a fluorescent imaging to show positive differentiation with this cell line? Or are there any other suggestions?
Edit: I am looking for a more quantifiable analysis of showing monocyte to macrophage differentiation with PMA treatment aside from looking at phenotypic responses in imaging.
Thanks so much
Hi Brianna,
You may use CD68, CD71 as markers for differentiated macrophages through immunofluorescence microscopy. You must withdraw PMA after initial incubation and let the cultures in media for at least 72 hours to have macrophages in their ground state.
Hi Briana
I agree with Rajeev, we usually use CD68 to check the differentiated of macrophage. we usually get the more than 80% differentiated macrophage after 48 hour with 50 nM PMA
You can perform real time PCR/western blotting for above said markers and determine the increase in these markers after differentiation with PMA.
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