Use of human AB serum or serum-derivatives for cell culture and manipulation steps comes with significant drawbacks and risk. Human AB serum contains undefined components, its performance can be influenced by donor-specific characteristics, and there is the potential to introduce pathogens into the workflow.

Consistent batch-to-batch product performance is necessary to develop standardized, controlled, and repeatable cell manufacturing workflow and processes. To take out the risk and inconsistency of serum in culturing cells, we offer serum-free solutions for the cell processing workflow. All Cell-Vive™ GMP cell culture products are manufactured with the highest quality raw materials, supplied by qualified vendors and supported by strict traceable documentation.

With us, you have a choice of formulations to suit different intended applications and experimental goals, while getting additional control, consistency, and safety for cell-based workflows.

Media Supplements for Immune Cells

Our Cell-Vive™ T-NK Xeno-Free Serum Substitute, GMP allows for a more controlled and consistent cell manufacturing process, leading to consistency in the expansion of T and NK cells.

Product Features:

PBMC-derived T cell culture was activated with anti-human CD3, anti-human CD28, and 200 IU/mL of recombinant human IL-2 using IMDM as basal media.

PBMC-derived NK cells were cultured using K-562 feeder cells (2:1) and 1000 IU/mL of recombinant human IL-2 using IMDM as basal media.

Cell-Vive™ T cell CD Serum Substitute, GMP is a chemically defined serum substitute additive made with non-animal derived components that supports T cell expansion and activation.

Product Features:

PBMC-derived T cells were activated with 1 µg/mL of Ultra-LEAF™ anti-human CD3 antibody (Cat. No. 317347), 1 µg/mL of Ultra-LEAF™ anti-human CD28 antibody (Cat. No. 302943) in IMDM basal media supplemented with 200 IU/mL Recombinant Human IL-2 (Cat. No. 791908) and media additives as indicated. On day 10, flow cytometry was used to determine specific T cell populations by making use of anti-human CD62L and anti-human CD45RO antibodies.

PBMC-derived T cells were activated with 1 µg/mL of Ultra-LEAF™ anti-human CD3 antibody (Cat. No. 317347), 1 µg/mL of Ultra-LEAF™ anti-human CD28 antibody (Cat. No. 302943) in IMDM basal media supplemented with 200 IU/mL Recombinant Human IL-2 (Cat. No. 791908) and media additives as indicated. (A) Cell expansion was determined on day 4, 6, 8, and end of culture (day 11). (B) Flow cytometry was used to determine specific T cell populations, on day 11, by anti-human CD4 (Cat. No. 317408) and anti-human CD8 (Cat. No. 344722) staining.

Our phenotypic analysis showed that the cells expanded in either the xeno-free or CD serum substitutes exhibited different functional properties. Using the same protocol, the xeno-free serum substitute tends to expand a greater proportion of naïve CD3⁺ T cell and regulatory T cell (Treg) populations, while the CD formulation preferentially expands central memory T cells and naïve T cells.

Human naïve CD4+ T cells were stimulated with Ultra-LEAF™ anti-human CD3 and Ultra-LEAF™ anti-human CD28 antibodies (1 µg/mL), and Recombinant Human IL-2 (200 IU/mL) alone (data not shown) or with the additions of Recombinant Human TGF-β (5 ng/mL) for 6 days in the presence of human AB serum, Cell-Vive™ T Cell CD Serum Substitute, GMP, or  Cell-Vive^TM T-NK Xeno-Free Serum Substitute. ANOVA was used to determine the effect of serum additives on Treg levels.*p<0.05

Media Supplements for Stem Cells

ITS (insulin, transferrin, sodium selenite) cell culture supplement contains these essential components for optimal viability and expansion of dozens of cell types. From iPSCs to many primary human neuronal cells, cardiomyocytes, and hepatocytes, ITS is used to support the viability and expansion of human cells of interest in the absence of FBS or other serum-based products. Although major cell culture reagent suppliers offer ITS supplement, our Cell-Vive™ CD ITS Media Supplement (100X), GMP is the first GMP-grade iteration of this cell culture supplement and is targeted towards the manufacturing of cell therapy products or for bioprocessing applications.

HeLa Cells were cultured in media containing EMEM and 10% fetal bovine serum for minimum 2-3 days. Cells were then split and seeded at a cell density of 3.5 x 10⁵ cells in a 6 well plate in EMEM 0.1% FBS and a final concentration of 1X of BioLegend Cell-Vive™ CD ITS Media Supplement, GMP (Cat. No. 420524, black bar) or a Competitor’s ITS solution (white bar) for 2-3 days. At the time of harvest, cells were dissociated with 0.25% trypsin EDTA and counted for (A) final cell counts and (B) viability.

Transplantation of neural stem cells (NSCs) has emerged as a promising treatment for neurological diseases due to the capacity of NSCs to secrete neurotrophic factors and promote nerve regeneration. We’ve designed Cell-Vive™ N-2 CD Prime Supplement (100X), GMP, for optimal expansion and differentiation of neural stem cells and neural progenitor cells. Its enriched formulation is chemically defined, animal component-free, and is precisely what’s needed for the preferential differentiation of NSCs. You will appreciate the consistent performance it offers when used in long term cultures, making it a critical component for ex vivo cell bioprocessing and manufacturing workflows.

Cell-Vive™ N-2 CD Prime Supplement (100X), GMP supports quality expansion of multipotent neural progenitor cells after expansion. Rat cortical stem cells (RCSCs) were grown for 7 days in vitro in media supplemented with Cell-Vive™ N-2 CD Prime Supplement (100X) in absence of recombinant human FGF-basic. Stem cell expression marker (Alexa Fluor® 594 anti-Nestin antibody, Cat. No. 655105, clone Rat-401) was used to stained undifferentiated RCSCs. Following the withdrawal of FGF basic, cells were randomly differentiated into neurons, astrocytes, and oligodendrocytes. Markers of lineage differentiation were detected using Alexa Fluor® 488 -conjugated anti-neuronal marker Tubulin β3 antibody (green, clone Tuj1, Cat. No. 801203) Alexa Fluor® 594 -conjugated astrocyte marker GFAP antibody (red, clone 2E1.E9, Ca. No. 644708) and Alexa Fluor 594®-conjugated oligodendrocyte marker O1 antibody (green, clone O1, Cat. No. 607755). Cell nuclei were counterstained with DAPI (blue). Round dot arrows indicate undifferentiated cells and solid arrows indicates differentiated cells.

Researchers can now select the media supplement that works best for their application and desired cell type. For any application inquiries you may have, please contact BioLegend Technical Support. Our team is ready to help.