B Cell Culture and Analysis

B cells represent a small portion of the peripheral blood lymphocyte population. Therefore, it is essential to expand B cells in vitro for many downstream applications, including functional studies, infectious disease research, and the development of vaccines and therapeutic antibodies.

Mouse B cells can be purified from splenocytes using magnetic bead-based negative selection, while human B cells are isolated from peripheral blood mononuclear cells by either negative selection, or CD19⁺ positive selection. B cells are then commonly cultured in vitro on a stromal feeder cell layer expressing CD40 Ligand in media supplemented with specific cytokines, which can include IL-4, IL-2, IL-21, and/or BAFF. Additional cytokines can be used to promote B cell differentiation, antibody production, and the generation of specific antibody isotypes such as IgA, IgG, IgE, or IgM.

Cytokines for B Cell Culture and Differentiation

R&D Systems large selection of cytokines can help optimize your B cell cultures. Our proteins are rigorously tested to ensure exceptional performance and lot-to-lot consistency, so you can have confidence in their ability to promote robust B cell expansion and differentiation with minimal variability between cultures. From basic research to clinical applications, we offer research-grade, Animal-free preclinical, and GMP-grade proteins to meet all research needs.

* GMP-grade proteins are available for these molecules.
† Liquid formulations of the GMP and Animal-free Preclinical grades of these proteins are available.

CellXVivo™ Human B Cell Expansion Kit

CellXVivo Immune Cell Expansion and Differentiation Kits contain optimized cytokine combinations and protocols for the expansion and differentiation of immune cell types. The CellXVivo Human B Cell Expansion Kit provides enough reagents for the 3-5-fold expansion of a starting population of 10⁷ human B cells.

ExCellerate™ Media for B Cell Expansion

ExCellerate B Cell Media is designed to promote robust B cell expansion, while ensuring consistent culture conditions. This media is specially formulated and optimized for the ex vivo culture of B cells under xeno-free conditions. It supports target antigen-specific B cell clonogenicity and the culture of mouse B cell hybridoma cell lines for antibody production, and it is compatible with B cells from multiple species.

Expansion of Mouse B Cells in ExCellerate™ Human B Cell Expansion Media Supplemented with Recombinant Mouse IL-4 and an Anti-Mouse CD40 Antibody. Mouse B cells were isolated from splenocytes using the MagCellect™ Mouse B Cell Isolation Kit (Catalog # MAGM204) and expanded using ExCellerate B Cell Media (Catalog # CCM031) with or without Recombinant Mouse IL-4 (Catalog # 404-ML) and a Rat Anti-Mouse CD40/TNFRSF5 Monoclonal Antibody (Catalog # MAB440). (A) Expansion of mouse B cells was monitored using Rezasurin (Catalog # AR002). Data show that ExCellerate B Cell Media supplemented with IL-4 and an Anti-Mouse CD40 Antibody results in robust mouse B cell expansion. (B) Representative brightfield images of mouse B cell colonies. (C) Expanded mouse B cells are B220⁺CD19⁺ (>98%) and negative for both CD3 and CD4.

Different B cell subsets are commonly identified based on the expression or lack of expression of specific cell surface and intracellular markers. Our selection of primary antibodies conjugated to one of over 30 fluorescent labels allows direct detection of cell surface and intracellular targets with the highest level of flexibility for multiplex experiments. Our antibodies are validated for specificity and reproducibility in multiple applications, including flow cytometry, immunocytochemistry/immunofluorescence (ICC/IF), immunohistochemistry (IHC), and Western blot. The tables below list the markers that are commonly used to identify different B cell subsets.

When an R&D Systems antibody is not available for a specific target molecule, we recommend using an antibody from our sister brand, Novus Biologicals™, whose products can be found at novusbio.com. Links provided in the tables below will show you to a list of antibodies from Novus Biologicals when an R&D Systems antibody is not available for a specific target.

Follicular B Cells

Follicular B cells are the largest subset of mature B cells in mice and humans and are found in the follicles of the lymph nodes and spleen. They recirculate in the secondary lymphoid organs and can differentiate into short-lived, antibody-secreting plasma cells and memory B cells following activation.

Marginal Zone B Cells

Marginal zone B cells are a subset of innate-like B cells located in between the white and red pulp in the marginal zone of the spleen, where they play a key role in protecting against blood borne pathogens. Marginal zone B cells have a poly-reactive BCR that can bind to a variety of antigens. Following activation, they produce low-affinity antibodies aimed at rapidly clearing pathogens and apoptotic debris.

Memory B Cells

Once B cells are activated, they differentiate into memory B cells in germinal centers that form within the B cell follicles in the lymph nodes and spleen. Memory B cells circulate in the blood and have BCRs that are specific to the antigens that triggered their initial formation. Upon re-exposure to the antigen, memory B cells quickly respond by producing high affinity, antigen-specific antibodies to protect the host from re-infection.

Plasma Cells

Plasma cells are antibody-secreting cells required for both short-lived and long-term antibody responses following antigen exposure. They are terminally-differentiated, non-proliferative cells, whose primary function is to secrete antibodies at a high rate to defend the host against an invading pathogen.

Regulatory B Cells

Regulatory B cells are a small subset of B cells with immunosuppressive properties that are involved in dampening inflammation and restoring immune homeostasis. These cells produce cytokines such as IL-10, IL-35, and TGF-β, which inhibit the differentiation, proliferation, and activation of pro-inflammatory T effector cells.

Detection of B Cells Using Multicolor Flow Cytometry. B cells were detected in human peripheral blood mononuclear cells (PBMCs) by staining with antibodies in the CD19 B Cell Panel (Catalog # FMC-P-003), which includes an Alexa Fluor^® 405-conjugated Mouse Anti-Human CD3 Monoclonal Antibody, an Alexa Fluor^® 647-conjugated Mouse Anti-Human CD45 Monoclonal Antibody, and a PE-conjugated Mouse Anti-Human CD19 Monoclonal Antibody. Alexa Fluor^® is a registered trademark of Molecular Probes, Inc., Eugene, OR.

Proteome Profiler™ Antibody Arrays

Learn more about Proteome Profiler Antibody Arrays.

Luminex^® Panels

Luminex panels are bead-based multiplex assays that enable you to multiplex up to 50 analytes, conserving on sample, time, and cost. Select from more than 490 targets spanning human, mouse, rat, porcine, and non-human primate species. Luminex Assays will deliver data that is reliable and reproducible because every panel is QC tested.

Learn more about Luminex Assays.

Luminex® is a registered trademark of Luminex Corporation.