I am studying phagocytosis/efferocytosis in vivo and have been using the peritoneum as a source of cells, but have a very low yield of dendritic cells. I also have a lot of CD19+ B-cells that take up 50-60% of the peritoneal exudate cells.
What would be the best tissue site for studying dendritic cell phagocytosis in vivo so that I can get a good yield.
Most protocols that use the peritoneum focus on macrophages.
I advise using fresh monocytes from whole blood separated by Ficoll. Then an antibody column to separate out murine monocytes. You can use rmuIL-4 and GM-CSF to turn them into 90% DC in 4-5 days.
Frozen cells don't work as well in these settings.
Bruce Lyday thanks for your response. However, I am doing the study in vivo so I won't be isolating monocytes to differentiate into DCs ex-vivo. What I need is which tissue site would be best to get already differentiated DCs in large numbers.
Skin would be best source IMHO, but harvesting might be challenging. DC make up
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