I isolated DCs from mice spleen using Miltenyi kit. The cells were provided with GM-CSF and IL4. Suspectedly, the adherent cells proliferate very robustly and behave almost like a cell line (already 3 wks in culture). An insight from anyone with an experience in DCs culture would be very appreciated.
Hi Yosef. Its been a long time since I produced dendritic cells and I've only done it from mouse bone marrow or human PBMC. 3 weeks seems like a long time for them to be in culture. When I did it, the DC were mature after 7 days and then I would use them for whatever assay I was doing at the time. Like I said, I never used spleens and I didn't use a kit, so maybe that makes it different.
I did culture of DCs from the bone marrow, where I differentiated monocytes into DCs with GM-CSF and IL-4 for 6 days. I never tried from spleen, but the 3 weeks culture is way too long for the cells. Is there a specific reason why you are targeting DCs from spleen?
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