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.

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.

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.

CellXVivo Human B Cell Expansion Kit

Name: CellXVivo Human B Cell Expansion Kit
Brand: R&D Systems
SKU: CDK005
Rating: 5 (1 reviews)

R&D Systems, part of Bio-Techne | Catalog # CDK005

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Citations (12)

Product Datasheet / COA / SDS

For the proliferative expansion of human B cell populations.

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.

Kit Contents

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.

Product Specifications for CellXVivo Human B Cell Expansion Kit

Species	Human
Source	N/A

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).

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.