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Key Benefits of Using R&D Systems Cytokines for Immune Cell Culture

  • High Levels of Biological Activity: Utilizing over 900 different bioassays, the biological activity of every protein we offer is tested in an appropriate bioassay to confirm that it meets our strict QC activity parameters.
  • Lot-to-Lot Consistency: Minimal lot-to-lot variability is ensured by testing each new lot side-by-side with previous lots and with a master lot, so you don’t have to worry whether results will be reproducible over time.
  • High Purity: Our proteins are typically over 95% pure.
  • Low Endotoxin Levels: Our proteins have a guaranteed industry-leading endotoxin level of <0.1 EU/ug by the LAL method.
  • Seamless Transition from Preclinical Research to Clinical Manufacturing: R&D Systems Animal-free RUO and Animal-free GMP-grade proteins frequently originate from the same clone, sequence, and expression system to make the transition from preclinical research into clinical manufacturing as seamless as possible.
  • Supply Chain Reliability: Our team has the experience and the capacity to ensure that we can provide you with a stable supply of the proteins you need for your research.
  • Bulk Proteins at Discounted Prices: We have the ability to scale up the production of any protein and we offer economical pricing on bulk orders.
  • Proteins Beyond the Catalog: We have tens of thousands of non-catalog proteins that may include different tags or come from different sources than the proteins listed on our website. Contact R&D Systems to see if we may already have the protein that you need.
  • Custom Protein Capabilities: For specialized protein requests, you can always contact our Custom Protein Services team. We have the expertise and the systems necessary to develop the proteins required to advance your research.
  • Comprehensive Portfolio of Reagents for Your Entire Workflow: Along with our proteins, Bio-Techne also offers a wide range of other products for immune cell culture and characterization, including media, media supplements, antibodies, immunoassays, RNAscope™ ISH assays, and analytical instruments to automate different steps of your workflow. 

T Cells

CD3+ T cells typically make up 70-85% of the lymphocyte population in healthy human adults and 45-70% of peripheral blood mononuclear cells. They are distinguished from other lymphocytes by their expression of the T cell receptor (TCR). CD8+ cytotoxic T lymphocytes (CTLs) are a subset of CD3+ T cells that recognize MHC class I- bound peptide ligands and directly kill damaged, infected, and dysfunctional cells. In contrast, CD4+ T cells are a subset of CD3+ T cells that recognize MHC class II-bound peptide antigens and play a central role in directing adaptive immune responses.

T cell

Cytokines for T Cell Culture or Differentiation - Proteins by Molecule

B7-H2/ICOS Ligand BMP-4* FGF-basic* Flt-3 Ligand/FLT3L* IFN-γ*
IL-1β* IL-2* IL-4* IL-6* IL-7*
IL-12 IL-15* IL-18/IL-1F4 IL-21* IL-23
IL-27 SCF* TGF-β1* TNF-α* Thrombopoietin*

*GMP-grade Proteins are available for these molecules.

Antibodies for T Cell Activation or Differentiation - Products by Molecule

CD3 CD28 IFN-γ IL-4 IL-12

 

Expansion of human T cells cultured for 5, 7, and 9 days in ExCellerate T Cell Expansion Media supplemented with IL-2, Detection of CD3+CD8+ and CD3+CD8- T cells by flow cytometry following enrichment in ExCellerate T Cell Expansion Media

Expansion of T Cells with R&D Systems RUO IL-2, Animal-Free RUO IL-2, and Animal-Free GMP-Grade IL-2 Proteins

 

PBMCs were isolated from 3 donors with the Fresenius Kabi Lovo® Automated Cell Processing System followed by positive selection of CD4+ and CD8+ T cells. Cells were expanded in a Wilson Wolf G-Rex® 6M Well Plate supplemented with 200 IU/mL of R&D Systems RUO Recombinant Human IL-2 (Catalog # BT-002), Animal-free RUO Recombinant Human IL-2 (Catalog # BT-002-AFL), or Animal-free GMP-grade Recombinant Human IL-2 (Catalog # BT-002-GMP) for 3 days, followed by 10-11 additional days with fresh medium and the same version of IL-2. Following this time, cells were analyzed for (A) expansion (B) viability, and (C, D) phenotype. Histogram bars represent the average of 3 donors. (C, D) Cells were analyzed by flow cytometry to determine the percentages of CD4+ or CD8+ T stem cell memory or central memory (Tscm/Tcm) cells compared with the percentages of effector or effector memory T cells (Teff/Tem) using antibodies against CD62L and CCR7. Little difference was observed in the CD4+ and CD8+ T cell phenotypes generated with the different IL-2 products.

*GMP-grade IL-2, IL-7, and IL-15, as well as G-Rex®, and Lovo® are available through our joint venture partnership with ScaleReady. G-Rex® is a registered trademark of Wilson Wolf Corporation. LOVO® is a registered trademark of Fresenius Kabi.

Natural Killer Cells

Natural killer cells are lymphocytes belonging to the innate immune system that account for 5-20% of the lymphocyte population and up to 15% of peripheral blood mononuclear cells. They have both intrinsic cytotoxic potential and cytokine-producing capabilities and as a result, they have been suggested to be the innate counterpart of CD8+ T cells. Human NK cells are phenotypically characterized as cells expressing high levels of CD56/NCAM-1 and lacking expression of CD3.

Natural Killer Cells

Cytokines for Natural Killer (NK) Cell Culture - Proteins by Molecule

BMP-4* Flt-3 Ligand/FLT3L* IFN-γ* IL-2* IL-3* IL-7*
IL-12 IL-15* IL-18/IL-1F4 IL-21* SCF* VEGF*

*GMP-grade Proteins are available for these molecules.

Expansion of Human Natural Killer Cells with ExCellerate Media and R&D Systems Cytokines

Expansion of human NK cells cultured in ExCellerate NK Cell Expansion Media with IL-2, IL-12, IL-18, and IL-21, Evaluation of CD3-CD56+ NK cell purity by flow cytometry following culture in ExCellerate NK Cell Expansion Media, Graph showing the purity of

Expansion of Human Natural Killer Cells in ExCellerate Human NK Cell Expansion Media Supplemented with Recombinant Human IL-2, IL-12, IL-18, and IL-21. Human peripheral blood mononuclear cells (PBMCs) were expanded for 14 days in ExCellerate Human NK Cell Expansion Media (R&D Systems, Catalog # CCM032) using NK cell-activating beads and Recombinant Human IL-2 (R&D Systems, Catalog # 202-IL), Recombinant Human IL-12 (R&D Systems, Catalog # 219-IL), Recombinant Human IL-18/IL-1F4 (R&D Systems, Catalog # 9124-IL), and Recombinant Human IL-21 (R&D Systems, Catalog # 8879-IL). (A) NK cell expansion was evaluated at days 0, 7, 9, and 14 under these culture conditions and showed an approximately 150-fold expansion following 14 days. (B) NK cell purity was also evaluated at days 0, 7, 9, and 14 by flow cytometry using a FITC-conjugated Mouse Anti-Human CD3 Monoclonal Antibody (R&D Systems, Catalog # FAB100F) and an APC-conjugated Mouse Anti-Human NCAM-1/CD56 Monoclonal Antibody (R&D Systems, Catalog # FAB2408A). Flow quadrants were set based on isotype control-stained samples. Following 14 days in culture, the CD3-CD56+ NK cell population showed a purity of 85.3% under these culture conditions as shown in the graph.

Monocytes/Macrophages

CD14+ monocytes are present in large numbers in the circulation and in the periphery, typically accounting for 10-20% of peripheral blood mononuclear cells in humans. Macrophages are derived from monocytes that differentiate to become inflammatory macrophages or tissue-resident macrophages, which are involved in maintaining tissue homeostasis and carrying out specialized tissue-specific functions, depending on their tissue of residence. Monocytes and macrophages both function as professional phagocytes that ingest and process foreign materials and cell debris. They also act as antigen-presenting cells and produce cytokines, allowing them to influence adaptive immune responses.

Monocytes / Macrophages

Cytokines for Monocyte/Macrophage Culture - Proteins by Molecule

GM-CSF* IFN-γ* IL-3* IL-4* IL-6*
IL-10* IL-13 M-CSF* SCF* Thrombopoietin*

*GMP-grade Proteins are available for these molecules.

Differentiation and Characterization of M1 and M2 Macrophages

Part A: 2 by flow cytometry indicating a CD14+CD80+CD163dimCD206+ phenotype. Part B:Characterization of differentiated M2 macrophages by flow cytometry indicating a CD14+CD80-CD163+CD206+ phenotype.

Detection of Cell Surface Markers on M1- or M2-Activated Human Macrophages by Flow Cytometry. Enriched human CD14+ monocytes were cultured in the presence of either (A) Recombinant Human GM-CSF (R&D Systems, Catalog # 215-GM) or (B) Recombinant Human M-CSF (R&D Systems, Catalog # 216-MC) for 6 days in serum-free base media to promote the differentiation of M1 or M2 macrophages, respectively. On day 6 of the differentiation, cells were harvested and stained with a Fluorescein-conjugated Mouse Anti-Human CD14 Monoclonal Antibody (open histogram; R&D Systems, Catalog # FAB3832F), a PE-conjugated Mouse Anti-Human B7-1/CD80 Monoclonal Antibody (open histogram; R&D Systems, Catalog # FAB140P), a PerCP-conjugated Mouse Anti-Human CD163 Monoclonal Antibody (open histogram; R&D Systems, Catalog # FAB1607C) and an APC-conjugated Mouse Anti-Human CD206/MMR Monoclonal Antibody (open histogram; R&D Systems, Catalog # FAB25342A). Cell staining was gated using isotype control antibodies (filled histograms). (A) M1 macrophages display a CD14+CD80+CD163dimCD206+ phenotype while (B) M2 macrophages display a CD14+CD80-CD163+CD206+phenotype.

Part A: ELISA results demonstrate that differentiated LPS-stimulated M1 macrophages secrete IL-12, while M2 macrophages do not.  Part B: ELISA results demonstrate that differentiated LPS-stimulated M2 macrophages secrete IL-10, while M1 macrophages do not

Differentiated Human M1 Macrophages Secrete IL-12, while Differentiated M2 Macrophages Secrete IL-10.Enriched human CD14+ monocytes were cultured in the presence of either (A) Recombinant Human GM-CSF (R&D Systems, Catalog # 215-GM) or (B) Recombinant Human M-CSF (R&D Systems, Catalog # 216-MC) for 6 days in serum-free base media to promote the differentiation of M1 or M2 macrophages, respectively. On day six, M1 and M2 macrophages were stimulated with 1 ug/mL LPS for 24 hours. Cell culture supernatants were collected and cytokine secretion was measured using the Human IL-12 p70 Quantikine™ HS ELISA Kit (R&D Systems, Catalog # HS120) and the Human IL-10 Quantikine™ ELISA Kit (R&D Systems, Catalog # D1000B).

Dendritic Cells

Dendritic cells are a rare cell population, accounting for only 1-2% of peripheral blood mononuclear cells in healthy humans, but they are key mediators of both innate and adaptive immune responses. They upregulate MHC molecules and costimulatory receptors upon pathogen recognition, capture, process and present antigens to naïve T cells, and produce polarizing cytokines that promote pathogen-specific effector T cell differentiation and activation. Dendritic cells are a heterogeneous population in terms of locations, phenotypes, and immunological functions, which allows them to differentially shape the immune response when presented with diverse pathogens. Multiple human and mouse dendritic cell subsets have been characterized.

Dendritic Cells

Cytokines for Dendritic Cell Culture - Proteins by Molecule

BMP-4* Flt-3 Ligand/FLT3L* GM-CSF* IFN-γ* IL-1β* IL-3*
IL-4* M-CSF* SCF* Thrombopoietin* TNF-α* VEGF*

*GMP-grade Proteins are available for these molecules.

Brightfield image of LPS-matured monocyte-derived dendritic cells following differentiation from CD14+ monocytes.

Differentiation and Characterization of Monocyte-Derived Dendritic Cells

 

Differentiation of CD14+ Monocytes into LPS-matured Monocyte-derived Dendritic Cells. CD14+ enriched peripheral blood mononuclear cells (PBMCs) were cultured for seven days in StemXVivo™ Serum-Free Dendritic Cell Base Media (R&D Systems, Catalog # CCM003) supplemented with Recombinant Human IL-4 (R&D Systems, Catalog # 204-IL) and Recombinant Human GM-CSF (R&D Systems, Catalog # 215-GM) and then induced with LPS for an additional 48 hours. Representative brightfield images of the cells on day 9 are shown.

Phenotypic Analysis of Cultured Monocyte-derived Dendritic Cells Before and After LPS-Induced Maturation.Immature monocyte-derived dendritic cells (open histograms, dotted black line) were obtained after CD14+-enriched monocytes were cultured for seven days in StemXVivo Serum-Free Dendritic Cell Base Media (R&D Systems, Catalog # CCM003) supplemented with Recombinant Human IL-4 (R&D Systems, Catalog # 204-IL) and Recombinant Human GM-CSF (R&D Systems, Catalog # 215-GM). Mature monocyte-derived dendritic cells (filled histograms) were obtained by culturing CD14+-enriched monocytes under the same conditions for seven days and then treating the cells with LPS for an additional 48 hours. The phenotypes of Day 7 immature monocyte-derived dendritic cells and day nine LPS-treated mature monocyte-derived dendritic cells were assessed by staining with a Fluorescein-conjugated Mouse Anti-Human B7-1/CD80 Monoclonal Antibody (R&D Systems, Catalog # FAB140F), a Fluorescein-conjugated Mouse Anti-Human CD83 Monoclonal Antibody (R&D Systems, Catalog # FAB1774F), a Fluorescein-conjugated Mouse Anti-Human B7-2/CD86 Monoclonal Antibody (R&D Systems, Catalog # FAB141F), and an Anti-MHC Class II Antibody, or an appropriate isotype control antibody (open histograms, solid blue line), followed by flow cytometry.

Analysis of immature MoDCs and LPS-treated mature MoDCs by flow cytometry with B7-1, CD83, B7-2, and MHC antibodies

Mature Dendritic Cells Induce Proliferation of Allogeneic T Cells. CD14+ monocytes were cultured for seven days in StemXVivo Serum-Free Dendritic Cell Base Media (R&D Systems, Catalog # CCM003) supplemented with Recombinant Human IL-4 (R&D Systems, Catalog # 204-IL), Recombinant Human GM-CSF (R&D Systems, Catalog # 215-GM) and gentamycin. The cells were subsequently treated with LPS for an additional 48 hours to induce dendritic cell maturation. Graded doses of mature monocyte-derived dendritic cells were incubated with 1 x 105autologous or allogeneic CD3+ T cells for five days. 3H-thymidine (3H-TdR) was added to the culture for the final 18 hours and T cell proliferation was measured using a scintillation counter. Results are presented as the mean cpm obtained from three experiments.

Assay demonstrating that graded doses of LPS-treated mature MoDCs stimulate proliferation of allogeneic CD3+ T cells

B Cells

B cells represent around 5-20% of the lymphocyte population in humans and up to 15% of peripheral blood mononuclear cells. They are distinguished from other lymphocytes by their expression of the B cell receptor (BCR). B cells play a key role in the adaptive immune response as they function as antigen-presenting cells, secrete regulatory cytokines, produce antibodies, and generate immunological memory. There are multiple subsets of B cells in both humans and mice, including naïve B cells, transitional B cells, memory B cells, plasma cells, and regulatory B cells, which can be distinguished based on the expression of different markers. The cell surface marker, CD19 is the most common marker used to identify B cells, as it is stably expressed throughout B cell differentiation until the plasmablast stage.

B Cells

Cytokines for B Cell Culture - Proteins by Molecule

BAFF/BLyS/TNFSF13B CD40 Ligand Flt-3 Ligand/FLT3L* IFN-γ* IL-2*
IL-4* IL-7* IL-10* IL-21*  

*GMP-grade Proteins are available for these molecules.

Graph showing robust expansion of mouse B cells in ExCellerate media with IL-4 and Anti-Mouse CD40 Antibody, Brightfield image of mouse B cell colonies expanded in ExCellerate media with IL-4 and Anti-Mouse CD40 Antibody, Flow cytometry analysis of mouse

Expansion of Mouse B Cells with ExCellerate Media and R&D Systems Cytokines

 

Expansion of Mouse B Cells in ExCellerate Human B Cell Expansion Media Supplemented with Recombinant Mouse IL-4 and an Anti-Mouse CD40 Antibody. Mouse B cells were isolated from splenocytes using the MagCellect™ Mouse B Cell Isolation Kit (R&D Systems, Catalog # MAGM204) and expanded using ExCellerate B Cell Media (R&D Systems, Catalog # CCM031) with or without Recombinant Mouse IL-4 (R&D Systems, Catalog # 404-ML) and a Rat Anti-Mouse CD40/TNFRSF5 Monoclonal Antibody (R&D Systems, Catalog # MAB440). (A) Expansion of mouse B cells was monitored using Rezasurin. Data show that ExCellerate B Cell Media supplemented with IL-4 and an Anti-Mouse CD40 Antibody 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.

Granulocytes

Granulocytes are polymorphonuclear leukocytes with small cytoplasmic granules that release enzymes to defend the host against invading pathogens. Granulocytes include basophils, eosinophils, mast cells and neutrophils. All of these are terminally differentiated cells that develop in the bone marrow and function primarily as part of the innate immune response, although they may also have antigen-presenting capabilities.

Granulocytes

Cytokines for Granulocyte Culture - Proteins by Molecule

G-CSF GM-CSF* IL-3* IL-5 IL-6*
M-CSF* SCF* Thrombopoietin* VEGF*  

*GMP-grade Proteins are available for these molecules.

TPO Graph

R&D Systems Cytokines Are Frequently More Active than Leading Competitors' Proteins

 

R&D Systems Recombinant Human Thrombopoietin/TPO Displays Higher Activity than Leading Competitors’ Thrombopoietin Proteins. The bioactivity of R&D Systems Recombinant Human Thrombopoietin (Catalog # 288-TPE; orange line) or recombinant human Thrombopoietin/TPO from two different competitors (blue and green lines) was assessed by measuring the ability of the proteins to stimulate proliferation of the MO7e human megakaryocytic leukemic cell line. The ED50 for this effect for R&D Systems Recombinant Human Thrombopoietin is 0.05-0.5 ng/mL, which is over 2-fold more active than the two competitors’ proteins.

Recombinant Human SCF/c-Kit Ligand Graph

R&D Systems Cytokines Are Rigorously Tested to Ensure Lot-to-Lot Consistency

 

Lot-to-Lot Consistency Testing of Recombinant Human SCF/c-kit Ligand. Two independent lots of Recombinant Human SCF/c-kit Ligand (R&D Systems, Catalog # 11010-SC) were tested for their ability to stimulate the proliferation of TF-1 human erythroleukemic cells. The ED50 for this effect is 1-5 ng/mL. Each trace on the graph represents Recombinant Human SCF/c-kit Ligand from a different manufacturing run, demonstrating the lot-to-lot consistency of the protein.

Additional Immune Cell-Related Products

Cell Culture Reagents

Cell Culture Reagents

Browse our complete collection of products for cell culture. From our ExCellerate™ T Cell, B Cell, or NK Cell Expansion Media and media supplements, to our FBS and CellXVivo™ Immune Cell Differentiation and Expansion Kits, we offer a comprehensive range of reagents to support immune cell expansion and differentiation.

adherent cells supported by matrix proteins

Serum-Free and Animal-Free Cell Culture Media

The elimination of serum and animal components in culture media is an important step towards a defined cell culture process and will also improve the safety profile of your cell product and simplify regulatory pathways. We offer serum-free, animal component-free media and media supplements to enhance the consistency of your cultures.

Flow Cytometry eHandbook Teaser

Flow Cytometry-Validated Antibodies for Identifying Immune Cell Types

Bio-Techne offers an unparalleled selection of fluorochrome-conjugated R&D Systems™ and Novus Biologicals™ flow cytometry-qualified antibodies for identifying or characterizing different immune cell types in mixed population samples. Choose from an expansive collection of both proprietary antibodies and some of the most highly referenced antibody clones on the market.

Fluorescent Dyes Probes

Fluorescent Dyes

Through the Tocris brand, Bio-techne offers a range of ready-to-conjugate dyes, from traditional small organic dyes to exclusive new technology, providing scientists with a full spectrum of choices for biomolecule conjugation and application.

cytokine inflammation

Immunoassays for Cytokine Detection

From our complete, ready-to-use Quantikine™ ELISA Kits to our multiplex Luminex® Assays and fully automated Simple Plex™ Assays, you can count on our immunoassays to deliver accurate, reproducible, high-quality data for every experimental sample that you test.

Immune Cell Therapy-Related Products

GMP Proteins Vial

Preclinical Animal-Free™ RUO and GMP-Grade Recombinant Proteins

Bio-Techne offers both R&D Systems Animal-free RUO Proteins and Animal-free GMP-grade Proteins, which are manufactured using identical expression systems and purification methods to ensure a seamless transition for researchers moving from preclinical research into clinical manufacturing.

Fluorokine App Note

Fluorokines™ Fluorescent-Labeled Proteins

Utilize our wide selection of fluorescent-labeled recombinant proteins to evaluate the expression of specific chimeric antigen receptors (CARs) on CAR-T or CAR-NK cells. Fluorokines are highly specific and allow CAR-expressing cells to be directly stained and detected by flow cytometry.

Organoid and 3-D Cell Culture Teaser

Organoid and 3-D Cell Culture Products

Co-culture of immune cells with tumor organoids is being increasingly utilized to study interactions that may occur in tumor microenvironments and to evaluate potential immunotherapies. Bio-Techne offers a complete collection of organoid culture reagents that are designed to promote robust organoid growth, while maintaining consistent, reproducible culture conditions.

Ella instrument

Analytical Solutions for Cell Therapies

Rigorously characterize your mid-process and final cell therapy products to ensure the safest possible therapies. Our analytical platforms and assays enable rapid and accurate analysis of cells, viral vectors, molecules, and contaminants.