Plasma Cell Isolation Kit II ( #130-093-628 )

Plasma Cell Isolation Kit II

Overview

The Plasma Cell Isolation Kit II has been developed for the isolation of plasma cells from human PBMCs or bone marrow.

Details

Background information
Human plasma cells are a heterogenous cell population comprising several subsets from short-lived and proliferative plasmablasts in lymphoid tissues, transitional plasma cells in peripheral blood to long-lived and non-dividing plasma cells in bone marrow^1–3. Plasma cells of all differentiation stages are identified by the expression of high levels of the CD38 antigen. Other surface markers, however, are differentially regulated dependent on the stage of differentiation and the spatial localization. Plasma cells in the blood circulation lack the expression of the typical B cell marker CD22 and were found to express lower levels of CD19 than mature B cells.¹ They are further characterized as being CD27⁺⁺, CD31⁺, CD44⁺, CD45⁺, CD56^–, CD62L⁺, CD86⁺, and HLA-DR⁺.^1,4 A subset of CD38⁺ blood plasma cells further expresses the CD138 antigen, whereas all CD38⁺ plasma cells are also CD138⁺ in bone marrow.⁴ For the isolation of malignant plasma cells from bone marrow, the addition of CD56 MicroBeads may be left aside to avoid depletion of CD56-expressing tumor cells.

Detailed separation procedure
Non-plasma cells are indirectly magnetically labeled with a cocktail of biotin-conjugated monoclonal antibodies and Anti-Biotin MicroBeads. The labeled cells are subsequently depleted by separation over a MACS Column. In the second step, pre-enriched plasma cells (flow-through fraction) are directly labeled with CD38 MicroBeads and isolated by positive selection from the pre-enriched plasma cell fraction.

Downstream applications
Isolated CD38⁺ plasma cells can be used for the analysis of signal transduction pathways and molecular analysis, for gene expression profiling, studies on plasma cell dysfunctions (in allergy, asthma, autoimmunity, or infectious diseases) or studies on the migrational behavior of plasma cells.

Columns

For depletion: LD or autoMACS® Columns. For positive selection: MS or autoMACS® Columns.

Figure 1

CD38⁺ plasma cells were isolated from human bone marrow using the Plasma Cell Isolation Kit II, an LD Column and a MidiMACS^™ Separator, and two MS Columns and a MiniMACS^™ Separator. Aliquots of the different cell fractions were fluorescently stained with CD19-APC and CD38-FITC for identification of plasma cells.

PBMCs before separation

Pre-enriched plasma cells after depletion of non-plasma cells

Enriched CD38⁺ plasma cells