CD138+ Plasma Cell Isolation Kit ( #130-092-530 )

CD138⁺ Plasma Cell Isolation Kit

Overview

The CD138⁺ Plasma Cell Isolation Kit was developed for the isolation of CD138⁺ CD45R(B220)^low/– CD19^low/– antibody-secreting plasma cells¹from spleen, lymph nodes, and bone marrow.

Details

Background information
CD138 (Syndecan-1) is a heparan sulfate-rich integral membrane proteoglycan, which functions as a matrix receptor for interstitial collagens, fibronectin, and thrombospondin. CD138 is predominantly expressed on the surface of epithelial cells in mature mouse tissues.² Its expression on cells of the B cell lineage correlates with their developmental stage, location, and adhesion. In mice, CD138 is expressed on pre-B and immature B lymphocytes in the bone marrow. It is lost when B cells emigrate into the periphery, is absent on circulating and peripheral B cells, and is re-expressed upon B cell differentiation into plasmablasts and plasma cells.³

Detailed separation procedure
The isolation of mouse plasma cells is performed in a two-step procedure. First, non-plasma cells are indirectly magnetically labeled with a cocktail of biotin-conjugated antibodies and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted by separation over a MACS® Column. In the second step, plasma cells are directly labeled with CD138 MicroBeads and isolated by positive selection from the pre-enriched cell fraction. The magnetically labeled plasma cells are retained on the column and eluted after removal of the column from the magnetic field. To achieve highest purities, the positively selected cell fraction, containing the plasma cells, is separated over a second column.

Downstream applications
CD138⁺ plasma cells isolated with the CD138⁺ Plasma Cell Isolation Kit can be used to analyze molecular pathways of signal transduction or gene expression profiling. Furthermore, they are suitable for studies on plasma cell dysfunctions, e.g. in allergy, asthma, autoimmunity, or infectious diseases and migrational behaviour of plasma cells.

Columns

For the first magnetic separation (depletion): LD or autoMACS® Columns; for the second magnetic separation (positive selection): MS or autoMACS Columns.

Figure 1

Plasma cells were isolated from a mouse spleen cell suspension by using the CD138⁺Plasma Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator. The cells were fluorescently stained with CD45R (B220)-APC, CD19-FITC, and CD138-PE. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Before separation

Plasma cells were isolated from a mouse spleen cell suspension by using the CD138<sup>+ </sup>Plasma Cell Isolation Kit, an LD and two MS Columns, a MidiMACS™ and a MiniMACS™ Separator. The cells were fluorescently stained with CD45R (B220)-APC, CD19-FITC, and CD138-PE. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Pre-enriched plasma cells after depletion of CD49b (DX5)⁺ cells and CD45R (B220)⁺ cells

Isolated CD138⁺ plasma cells

Figure 2

Plasma cells were isolated from a mouse bone marrow cell suspension by using the CD138⁺ Plasma Cell Isolation Kit. The CD138⁺ plasma cells were stained intracellularly with goat anti-mouse IgG-PE by applying the solid-phase intracellular staining procedure (Inside Stain Kit, # 130-090-477). Additionally, the cell surface of the CD138⁺ plasma cells was fluorescently stained with CD138-APC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

Plasma cells were isolated from a mouse bone marrow cell suspension by using the CD138<sup>+</sup> Plasma Cell Isolation Kit. The CD138<sup>+</sup> plasma cells were stained intracellularly with goat anti-mouse IgG-PE by applying the solid-phase intracellular staining procedure (Inside Stain Kit, # 130-090-477). Additionally, the cell surface of the CD138<sup>+</sup> plasma cells was fluorescently stained with CD138-APC. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.