CellXVivo Human B Cell Expansion Kit
CellXVivo Human B Cell Expansion Kit Summary
For the proliferative expansion of B cell populations.
Key Benefits
- Yields a 3-5 fold expanded population of B cells
- Provides optimized reagents needed to induce B cell proliferation
- Utilizes validated and straightforward procedures
- Does not require specialized instrumentation
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.
Specifications
Product Datasheets
Scientific Data
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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.
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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).
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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
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
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
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.
Citations for CellXVivo Human B Cell Expansion Kit
R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.
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Citations: Showing 1 - 7
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Targeting CD74 in B-cell non-Hodgkin lymphoma with the antibody-drug conjugate STRO-001
Authors: X Li, C Abrahams, A Yu, M Embry, R Henningsen, V DeAlmeida, S Matheny, T Kline, A Yam, R Stafford, T Hallam, M Lupher, A Molina
Oncotarget, 2023-01-12;14(0):1-13. 2023-01-12
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The Rationale for the Dual-Targeting Therapy for RSK2 and AKT in Multiple Myeloma
Authors: R Isa, M Horinaka, T Tsukamoto, K Mizuhara, Y Fujibayash, Y Taminishi-, H Okamoto, S Yasuda, Y Kawaji-Kan, Y Matsumura-, S Mizutani, Y Shimura, M Taniwaki, T Sakai, J Kuroda
International Journal of Molecular Sciences, 2022-03-08;23(6):. 2022-03-08
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Tumor-B-cell interactions promote isotype switching to an immunosuppressive IgG4 antibody response through upregulation of IL-10 in triple negative breast cancers
Authors: NJ Toney, LM Opdenaker, K Cicek, L Frerichs, CR Kennington, S Oberly, H Archinal, R Somasundar, J Sims-Mourt
Journal of Translational Medicine, 2022-03-07;20(1):112. 2022-03-07
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IgA transcytosis and antigen recognition govern ovarian cancer immunity
Authors: S Biswas, G Mandal, KK Payne, CM Anadon, CD Gatenbee, RA Chaurio, TL Costich, C Moran, CM Harro, KE Rigolizzo, JA Mine, J Trillo-Tin, N Sasamoto, KL Terry, D Marchion, A Buras, RM Wenham, X Yu, MK Townsend, SS Tworoger, PC Rodriguez, AR Anderson, JR Conejo-Gar
Nature, 2021-02-03;0(0):. 2021-02-03
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B cell-specific XIST complex enforces X-inactivation and restrains atypical B cells
Authors: B Yu, Y Qi, R Li, Q Shi, A Satpathy, HY Chang
bioRxiv : the preprint server for biology, 2021-01-05;0(0):. 2021-01-05
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BAFF-driven NLRP3 inflammasome activation in B cells
Authors: KH Lim, LC Chen, K Hsu, CC Chang, CY Chang, CW Kao, YF Chang, MC Chang, CG Chen
Cell Death Dis, 2020-10-01;11(9):820. 2020-10-01
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A DNA Vaccine Protects Human Immune Cells against Zika Virus Infection in Humanized Mice
Authors: G Yi, X Xu, S Abraham, S Petersen, H Guo, N Ortega, P Shankar, N Manjunath
EBioMedicine, 2017-10-06;25(0):87-94. 2017-10-06
FAQs
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Can the CellXVivo Human B Cell Expansion Kit, catalog # CDK005, be used for expansion of memory B cells?
Based on the components used in the CDK005 kit, the kit is expected to expand total B cells and is not specialized for memory B cells.
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Will ExCellerate B Cell Media, Xeno-Free, Catalog # CCM031, promote expansion of B cell clones without addition of growth factors?
Catalog # CCM031 must be supplemented with growth factors in order to use it for expansion of B cell clones. Information on appropriate growth factor cocktails are available in published literature. Here are two references:
1. LeBien, T.W. and T.F. Tedder. (2008) Blood 112:1570.
2. Basso, K. and R. Dalla-Favera (2015) Nat. Rev. Immunol. 15:172.
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Can CellXVivo Human B cells expansion kit, Catalog # CDK005, also promote B cell Maturation (differentiation into plasma cells)?
Catalog # CDK005 has only been tested for B cell expansion. This kit has not been tested for B cell Maturation.
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