CD11c Microbeads, Mouse Miltenyi Biotech

Miltenyi Biotech manufactures several cell isolation systems.  These systems are based on magnetic separation.  Antibodies conjugated to magnetic beads are used to label cells; depending on the antibody, the beads will result in either positive or negative selection of a specific cell type.  In the case of the CD11c Microbeads, mouse dendritic cells in single cell suspensions derived from spleen, lymph nodes, Peyer’s patches, colonic lamina propria, bone marrow, epidermis, lung, liver, or brain will be positively selected and isolated.  Once isolated, these cells are useful for phenotypic and functional assays. Keep in mind, these beads can be used to deplete dendritic cells from a cell suspension.

To isolate dendritic cells using the CD11c Microbeads is quite simple. Essentially, one must first obtain of a single cell suspension (see Miltenyi’s webpage for suggested protocols) and to that suspension you will have to apply an appropriate amount of the bead suspension (based on number of total cells).  Incubate bead/cell suspension in the refrigerator (not on ice, it does make a difference) for 15 minutes, wash and run suspension through a magnetic column, remove the column from the magnet and collect the magnetically labeled cells. 

In my hands, these beads always produce reproducibly pure dendritic cells. This system is based on positive selection: the flow-through fraction of cells (non-dendritic cells) are unlabeled, subsequent cell isolation can be performed.  I commonly purify dendritic cells and then subsequently purify Gamma Delta T cells from intestinal tissue. The back-to-back cell isolation serves to save precious samples.  Since the dendritic cell isolation can be accomplished in a short amount of time, I can do multiple isolations without subjecting myself to an overwhelming and long workday. Important to note, in addition to following the manufacture’s instructions, I would strongly suggest the use of density gradient separation to remove the dead cells as they may bind to antibodies non-specifically.  Also I would recommend the use of the FcR Blocker prior to cell purification techniques to reduce non-specific interactions. 

One major issue when using Miltenyi’s systems for cell isolation is that the upfront cost for separation columns, magnet and the microbeads.  Although costly, Miltenyi is well known for their isolation systems. For preliminary experiments, you may want to use another investigators system. Also, since Miltenyi has such a huge presence in cell isolation systems, it is likely you could find columns and/or a magnet in a neighboring lab.  Furthermore, contact you Miltenyi sales representative they can be quite accommodating.