I am currently trying to isolate macrophages from pbmc using a Miltenyi kit based on magnetic beads. However, when I culture CD14+ cells, they dead rapidly. I think that macrophages are catching that beads and maybe it is the cause of their death. Has anyone more information about this? Thank you
Dear Elisa,
we also isolate macrophages with a magnetic bead system from Milteny (peritoneal macrophages with CD11b though) and we never encountered macrophage death after isolation and subsequent cultivation. So I think it is very unlikely that the magnetic beads themselves are the reason for your observed cell death. Perhaps you can give some more details about the sorting procedure?
All the best and stay healthy,
Marc
CD14 cells are the monocytes not the macrophages. They are very much sensitive to the stress and differentiate. Usually, monocytes dont stay in culture until provided specific growth factors to be differentiated into any lineage.
Possible reason for the CD14+ monocytes's death is mechanical stress and non availability of specific growth factors.
Dear Elisa Arribas , your CD14 cells are basically Monocytes, which may get activated due to stress in the separation process and died. Monocytes gets activated very soon and they are terminally differentiated cell so will not sustain in culture (proliferate), they will die sooner (unless they are cell lines). They may be differentiated in macrophages or DC by supplementing with specific growth factors.
Death in your separation or culture may be stress induced.