Plasma cell research

  • Isolation and culture of plasma cells from whole blood and bone marrow
  • Expansion of plasma cells in vitro
  • Analysis of subsets in flow cytometry

Application data for plasma cell research

Plasma cell separation

Magnetic Plasma cell separation with MACS® Technology

Generally, plasma cells have a very rare distribution in peripheral blood and are difficult to obtain from human bone marrow samples, which makes them particularly difficult to separate. Whether you require, purified populations separated by positive selection, two step protocols or separation directly from whole blood or bone marrow, MACS Technology has a solution for you.

Plasma cell isolation directly from blood products

StraightFrom® MicroBeads allow magnetic cell separation directly from human blood products, such as whole blood, buffy coat, leukoreduction system chambers (LRSCs) and leukopaks. CD19+ and CD138+ B cells can be purified using the autoMACS® Pro Separator, the MultiMACS™ Cell24 Separator Plus, or manual separators. For Plasma cell separations two products are available StraightFrom Whole Blood and Bone Marrow CD138 MicroBeads and StraightFrom® Whole Blood CD138 MicroBeads. For processing bone marrow samples we recommend our StraightFrom® Whole Blood and Bone Marrow CD138 MicroBeads to ensure the best performance.
 

StraightFrom Whole Blood and Bone Marrow CD138 MicroBeads

Separation of CD138+ cells from a whole blood sample spiked with U266 cells, using the StraightFrom® Whole Blood and Bone Marrow CD138 MicroBeads. Cells were stained with CD45-FITC, CD138-PE, and CD19-APC and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris, dead cells, and CD45 cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.
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Application protocol
Saving valuable hands-on time – automated CD138+ plasma cell enrichment

Plasma cell culture

Plasma cells are present in peripheral blood in very low frequencies and are difficult to obtain from human bone marrow samples. Therefore, in vitro expansion is an ideal choice to increase plasma cell quantities; either to study their biology or analyze their antibody production and specificity.  

We have developed the B Cell Expansion Kit, human, for easy and enhanced expansion. The formulation offers the perfect solution to expand and activate human B cells from various sources. This enables expansion of B cells that are fully functional and ready for any downstream application. The culturing conditions provided by the B Cell Expansion Kit and IL-21 drives B cell differentiation towards a matured, switched status. B cells cultured with the kit and IL-21, instead of IL-4, demonstrated a significant increase in the plasma cell fraction. This way you can expand the Plasma cell population before separating plasma cells for further experiments. 
 

A comparison of unstimulated B cells and those expanded with the B Cell Expansion Kit and the use of the B Cell Expansion Kit with IL-21 instead of IL-4

Comparison of unstimulated B cells and those expanded with the B Cell Expansion Kit 
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B cells were isolated on day 0 from human buffy coat using CD19 MicroBeads, human after performing a density gradient and cultured applying the low-density protocol provided with the B Cell Expansion Kit. One culture was cultured with addition of human IL-21 instead on human IL-4. At day 14, stimulated and unstimulated, showed a reduction of the naive B cell subset, in comparison to day 0, what is due to their described shorter survival rate. In the presence of IL-21, a significant increase in the plasma cell fraction (up to 20% of all B cells) was observed while switched memory B cells doubled from 22% to 45%. This suggests that the culturing condition provided by the B Cell Expansion Kit and IL-21 drives B cell differentiation towards a matured, switched status.

 

Plasma cell expansion rates

Reliable activation and expansion of Plasma cells is a definitive requirement for effective downstream experiments. The B Cell Expansion Kit, human is optimized for primary B cell expansion in vitro. With the use of human IL-21, premium grade a slight increase of the expansion rate can be detected on day 7 and 10.  

The B Cell Expansion Kit, human offers: 

  • Up to a 10-fold expansion of B cells in vitro within 14 days
  • Standardized and reproducible system 
     
B cell expansion rates using the B Cell Expansion Kit  
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B cell expansion rates using the B Cell Expansion Kit  

Applying the low-density protocol provided with the B Cell Expansion Kit an increase in the expansion rate of up to 10-fold could be achieved after 14 days of culture. A slight increase in expansion was already detected on days 7 and 10. This effect was further enhanced by replacement of IL-4 with IL-21. The modified procedure resulted in an earlier onset of expansion compared to the standard procedure of the B Cell Expansion Kit. To facilitate expansion, B cells were isolated on day 0 from human buffy coat using CD19 MicroBeads, human after performing a density gradient. 
 

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Scientific poster
A standardized and reproducible method for expansion and activation of human primary B cells 

Aparajita Singh,  Susanne Bethke, Daniela Vorholt, and Andrzej Dzionek 

Multiparameter flow analysis

As the final step in many workflows, flow analysis plays an important part in various B cell applications. The following flow reagents can help you make sure that the data you generate on your flow cytometer are reliable and consistent: 

  • REAfinity™ Recombinant Antibodies – recombinantly generated antibodies that ensure lot-to-lot consistency and eliminate background signal
  • MACSplex™ Cytokine Kits – analysis of multiple soluble cytokines in one sample using any standard flow cytometer
  • Cytokine Secretion Assay – Detection Kits – flow analysis of cytokine secretion at a detection level of one in a million
  • Build your own B cell flow panel with our Flow Panel Builder

Application protocol: Multicolor flow cytometric analysis of B cell subsets from mouse spleen

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