Differentiating Human Monocytes - (Jul/25/2005 )

I want to isolate monocytes from PBMC and differentiate them into dendritic cells. I've heard they require more than GMCSF/IL4. One person told me they need TNFa and phorbol ester. Does anyone have experince with this?

Thanks

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They don't need more than GM-CSF or IL-4 if you just plan to differentiate them to imDC. There are a number of protocals out there. We use the standard Ficoll gradient followed by adhesion to flasks and then GM-CSF. If you look up any human DC papers they should have this procedure. If not I can always send our lab's to you.

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and then
how can i detect those markers like CD83 , flow cytometry or others. extract mRNA for what
what can i do with the human peripheral blood, the protocol...
help me ...

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