Can ExCellerate B Cell Media, Xeno-Free, Catalog # CCM031, be used alone (without supplements) to keep B cells viable for a short period of time (24-48 hrs)?

B cells are expected to survive in the base medium for 24 to 48 hours, but without cytokine supplementation, there would be no cell proliferation and possibly some cell loss. We have not studied this extensively.

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 Excellerate B cell media, Xeno-free, Catalog # CCM031, be used for the expansion of Mouse B cells?

Yes. The Excellerate B cell media, Xeno-free, can be used for expansion of Mouse B cells. There are example data on our webpage in the Scientific data section.

Which mouse-specific reagents are needed when Catalog # CCM031 is used for expansion of murine B cells? What protocol should be followed?

Firstly, the mouse B cells should be isolated using Catalog # MAGM204 (MagCellect Mouse B Cell Isolation Kit) or equivalent. To prepare for B cell expansion, Excellerate B cell medium should be supplemented with recombiant mouse IL-4 protein and Mouse CD40/TNFRSF5 Antibody (Catalog # 404-ML and MAB440) respectively. A protocol for B cell expansion based on extensive published literature should be followed.

In the data shown for Mouse B cell expansion with Catalog # CCM031, was a cross-linking antibody used?

No cross-linking antibodies were used for Mouse B cell expansion.

ExCellerate B Cell Media, Xeno-Free Best Seller

Name: ExCellerate B Cell Media, Xeno-Free
Brand: R&D Systems
SKU: CCM031
Rating: 5 (1 reviews)

Catalog # CCM031 | R&D Systems, Inc. a Bio-Techne Brand

(1)

Product Datasheet / COA / SDS

Expansion and clonal growth of B cells.

Key Product Details

Setting the standard in quality research reagents

Technical Data
Citations / Reviews (1) / FAQs

Summary for ExCellerate B Cell Media, Xeno-Free

Summary

Media for the ex vivo culture of B lymphocytes under xeno-free conditions.

Key Benefits

Robust B cell expansion under xeno-free conditions
Facilitates the isolation and expansion of B cells from peripheral blood
Compatible for culturing B cells from multiple species
Supports target antigen-specific B cell clonogenicity
Supports culture of mouse B cell hybridoma cell lines for antibody production

Why Use ExCellerate B Cell Media?

B lymphocytes (B cells) play an essential role in the humoral component of the adaptive immune response. Following exposure to a pathogen or foreign material, B cells produce and secrete antibodies that circulate through the body and bind to specific pathogens, marking them for destruction. B cells also contribute to cellular immunity by maintaining immune homeostasis, including the regulation of T lymphocyte activation and expansion. Diseases or abnormalities that affect B cells have been shown to result in immunodeficiencies, autoimmunity, and cancer. Continued progress in basic and clinical B cell research is crucial for further understanding human health and disease.

ExCellerate B Cell Media is formulated and optimized for the ex vivo culture of B lymphocytes under xeno-free conditions. Unlike traditional serum-containing media, ExCellerate B Cell Media provides a stable and optimized culture environment, void of non-human animal-derived products, that facilitates the isolation and expansion of B cells from peripheral blood. This media supports target antigen-specific B cell clonogenicity as well as robust culture of mouse B cell hybridoma cell lines for antibody production.

Versatility of ExCellerate B Cell Media.

ExCellerate B Cell Media can be used with cytokine/growth factor supplements for the ex vivo culture of B cells. This versatile medium is compatible for culturing B cells from multiple species (i.e., human, mouse, rabbit, and canine). Cytokine/growth factor combination and concentrations should be used with this media should be optimized by application or experimental protocol.

Product Details

500 mL of ExCellerate B Cell Media, Xeno-Free

HSC Differentiation &Lineage Marker Schematic View Interactive Pathway

The definitive hematopoietic system is made up of all adult blood cell types including megakaryocytes, erythrocytes, and cells of the myeloid and lymphoid lineages. All of these cells are derived from multipotent hematopoietic stem cells (HSCs) through a succession of precursors with progressively limited potential. Hematopoietic stem cells are tissue-specific stem cells that exhibit remarkable self-renewal capacity and are responsible for the life-long maintenance of the hematopoietic system. HSCs are rare cells that reside in adult bone marrow where hematopoiesis is continuously taking place. They can also be found in cord blood, fetal liver, adult spleen, and peripheral blood. R&D Systems offers several products for studying hematopoietic lineage cells including serum-free media, lineage depletion antibodies and kits, and reagents for performing colony forming cell (CFC) assays.

Product Specifications for ExCellerate B Cell Media, Xeno-Free

Species

Human, Multi-Species

Scientific Data Examples for ExCellerate B Cell Media, Xeno-Free

	Improved B Cell Expansion using ExCellerate B Cell Media. Human B cells were isolated from peripheral blood mononuclear cells (PBMCs) and cultured for 10 days in either ExCellerate B Cell Media or RPMI + 10% FBS. Both media conditions were supplemented with reagents included in the CellXVivo Human B Cell Expansion Kit (Catalog # CDK005). A) Fold expansion of B cells at 5 and 10 days in culture (n=3). B) Light microscopy of B cells at 20X magnification after 5 days of culture.
	Detection of CD19 but not CD3 in Human B Cells. Human B Cells were cultured for 5 days in ExCellerate B Cell Media, supplemented with reagents included in the CellXVivo™ Human B Cell Expansion Kit (Catalog # CDK015). The cells were labeled with a Mouse Anti-Human CD19 PE-conjugated Antibody (Catalog # FAB4867P) and with a Mouse Anti-Human CD3 epsilon APC-conjugated (Catalog # FAB100A). CD19⁺CD3^- B cells were observed (>97%) following this culture protocol.
	Improved Positive Clone Isolation of B cells using ExCellerate B Cell Media. Rabbit B cells were isolated from PBMCs and plated into 384-well plates (1 cell/well) using either ExCellerate B Cell Media or RPMI + 10% FBS. Clonal growth was monitored and screening was performed to identify antigen-specific B cell clones using an ELISA-based protocol and with PCR-based monoclonal cloning of heavy and light chains. Positive identification of clones producing the correct antibody was >2.5 fold higher when cultured using ExCellerate B Cell Media compared to RPMI + 10% FBS (n=8).
	Expansion of Mouse B Cells using ExCellerate B Cell Media. Mouse B cells were isolated from splenocytes using the MagCellect Mouse B Cell Isolation Kit (Catalog # MAGM204) and expanded using ExCellerate B Cell Media with or without Recombinant Mouse IL-4 and CD40 Ab. A) Expansion of mouse B cells was monitored using Rezasurin. Data show that Excellerate B Cell Media supplemented with IL-4 (Catalog # 404-ML) and CD40 Ab (Catalog # MAB440) 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.

Preparation & Storage

Storage

Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Citations for ExCellerate B Cell Media, Xeno-Free

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FAQs for ExCellerate B Cell Media, Xeno-Free

Can ExCellerate B Cell Media, Xeno-Free, Catalog # CCM031, be used alone (without supplements) to keep B cells viable for a short period of time (24-48 hrs)?

B cells are expected to survive in the base medium for 24 to 48 hours, but without cytokine supplementation, there would be no cell proliferation and possibly some cell loss. We have not studied this extensively.
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 Excellerate B cell media, Xeno-free, Catalog # CCM031, be used for the expansion of Mouse B cells?

Yes. The Excellerate B cell media, Xeno-free, can be used for expansion of Mouse B cells. There are example data on our webpage in the Scientific data section.
Which mouse-specific reagents are needed when Catalog # CCM031 is used for expansion of murine B cells? What protocol should be followed?

Firstly, the mouse B cells should be isolated using Catalog # MAGM204 (MagCellect Mouse B Cell Isolation Kit) or equivalent. To prepare for B cell expansion, Excellerate B cell medium should be supplemented with recombiant mouse IL-4 protein and Mouse CD40/TNFRSF5 Antibody (Catalog # 404-ML and MAB440) respectively. A protocol for B cell expansion based on extensive published literature should be followed.
In the data shown for Mouse B cell expansion with Catalog # CCM031, was a cross-linking antibody used?

No cross-linking antibodies were used for Mouse B cell expansion.