CellXVivo Human B Cell Expansion Kit

For the proliferative expansion of B cell populations.

Why Expand B Cells Ex Vivo?

B cells are a subset of lymphocytes that express immunoglobulins on their cell surface. The B cell population is a vital component of the humoral immune response. B cell activation leads to antigen uptake and presentation, affinity maturation and class switching of the immunoglobulins, and antibody secretion. B cells represent a small portion (~5%) of the peripheral lymphocyte population making it difficult to obtain sufficient B cell quantities for many research applications. The ability to expand B cell populations ex vivo is valuable to researchers who require B cells for downstream applications including high throughput B cell assays and ex vivo differentiation of T cells.

This kit contains the following optimized reagents for the expansion of human B cell populations.

• B Cell Expander Reagents
• Reconstitution Buffers
• Provides enough reagents for the 3-5 fold expansion of a starting population of 10⁷ human B cells

Stability and Storage

Store the unopened kit at 2-8 ^°C. Do not use past the kit expiration date. Opened or reconstituted B Cell Expander 1, B Cell Expander 2, and B Cell Expander 3 should be stored at 2-8 ^°C under sterile conditions for up to 30 days or at -20 ^°C to -70 ^°C in a manual defrost freezer for up to 3 months.* Opened Reconstitution Buffer 1 and Reconstitution Buffer 2 should be stored at 2-8 ^°C under sterile conditions for up to 12 months.*

*Provided this is within the expiration date of the kit.

Scientific Data Examples for CellXVivo Human B Cell Expansion Kit

	Light Microscopy of B Cells. B cells were imaged at 20X magnification following isolation from PMBCs and (B) after 5 days in culture using reagents included in the Human B Cell Expansion Kit.
	Detection of CD20 in Human B Cells. Human B cells were expanded for 5 days using reagents included in the Human B Cell Expansion Kit. The cells were labeled with a PE-conjugated Mouse Anti-Human CD20 Monoclonal Antibody (R&D Systems, Catalog # FAB4225P; filled histogram) or a PE-conjugated Mouse IgG1 Isotype control (R&D Systems, Catalog # IC002P; open histogram).
	Human B Cell Expansion.  Human B cells were isolated from PMBCs and cultured for 5 days using reagents included in the Human B Cell Expansion Kit. B cell expansion was measured with Resazurin (R&D Systems, Catalog # AR002).

Assay Procedure

Refer to the product datasheet for complete product details.

 

Briefly, human B cell populations can be expanded using the following procedure:

• Isolate human B cells from PBMC preparation
• Culture B cells in B Cell Expander-supplemented media
• Measure B cell expansion using Resazurin

View Graphic Procedure

 

Reagents Provided

Reagents Supplied in the CellXVivo™ Human B Cell Expansion Kit (Catalog # CDK005):

• B Cell Expander 1
• B Cell Expander 2
• B Cell Expander 3
• Reconstitution Buffer 1
• Reconstitution Buffer 2

 

Other Supplies Required

Reagents

• MagCellect™ Human B Cell Isolation Kit (R&D Systems, Catalog # MAGH103, or equivalent).
• Resazurin (R&D Systems, Catalog # AR002, or equivalent).
• Ficoll-Hypaque™
• RPMI 1640
• L-Glutamine
• Penicillin
• Streptomycin
• Fetal Bovine Serum (FBS)
• beta-Mercaptoethanol

Equipment

• Tissue culture flasks and/or plates
• Inverted microscope
• Hemocytometer
• 37 °C and 5% CO₂ humidified cell culture incubator
• Benchtop centrifuge
• Pipettes and pipette tips

 

Procedure Overview

R&D Systems Protocol for Human B Cell expansion

Isolate PBMCs from human blood.

Isolate human B cells from a PBMC preparation (e.g., using magnetic cell selection).

Perform a cell count.

Culture 2 x 10⁵ cells/mL in media supplemented with reconstituted B Cell Expanders 1-3.

Incubate the cells for 5 days.

Measure B cell expansion using Resazurin.