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- CellXVivo Human B Cell Expansion Kit (CDK005)
CellXVivo Human B Cell Expansion Kit
R&D Systems, part of Bio-Techne | Catalog # CDK005
Key Product Details
Summary for CellXVivo Human B Cell Expansion Kit
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.
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.
| Species | Human |
| Source | N/A |
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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). |
Preparation & Storage
| Shipping Conditions | The product is shipped with polar packs. Upon receipt, store it immediately at the temperature recommended below. |
| Storage | Store the unopened product at 2 - 8 °C. Do not use past expiration date. |
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.