ExCellerate B Cell Media, Xeno-Free Summary
Media for the ex vivo culture of B lymphocytes under xeno-free conditions.
- 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.
- 500 mL of 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.
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.
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