YILIMART - Plasma Cell Isolation Kit II, human

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

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.

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They are further characterized as being CD27

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, CD31

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, CD44

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, CD45

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, CD56

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, CD62L

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, CD86

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, and HLA-DR

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.

^1,4

A subset of CD38

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blood plasma cells further expresses the CD138 antigen, whereas all CD38

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plasma cells are also CD138

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in bone marrow.

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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

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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 the first magnetic separation (depletion): LD or autoMACS

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Columns. For the second magnetic separation (positive selection): MS or autoMACS Columns.

Figure 1

CD38

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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.

Bone marrow before separation	Pre-enriched plasma cells after depletion of non-plasma cells

Enriched CD38+ plasma cells

Selected references
1. Medina et al. (2002) The heterogeneity shown by human plasma cells from tonsil, blood, and bone marrow reveals graded stages of increasing maturity, but local profiles of adhesion molecule expression. Blood 99: 2154-2161
2. Slifka, M. K. et al. (1998) Humoral immunity due to long-lived plasma cells. Immunity 8: 363-372
3. Manz, R. A. et al. (1997) Lifetime of plasma cells in the bone marrow. Nature 388: 133-134
4. Horst, A. et al. (2002)
  
  Detection and characterization of plasma cells in peripheral blood: correlation of IgE
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  plasma cell frequency with IgE serum titre.
  
  Clin. Exp. Immunol. 130: 370-378

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Plasma Cell Isolation Kit II, human

For research use only

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Background information

Detailed separation procedure

Downstream applications

Columns

Figure 1

Selected references

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