GM-CSF Reporter HEK 293 Cells

Cytokine offer: For each cytokine reporter cell line purchased, get a free vial of the matching cytokine. View all InvivoGen's Cytokine Bioassays.

HEK-Blue™ GM-CSF Cells signaling pathway

Granulocyte-Macrophage Colony-Stimulating Factor Reporter Cells

HEK-Blue™ GM-CSF cells are designed to monitor human GM-CSF-induced STAT5 stimulation or inhibition through SEAP detection. This colorimetric bioassay can be used for screening activatory molecules, such as engineered cytokines, or inhibitory molecules, such as neutralizing antibodies.

HEK-Blue™ GM-CSF cells respond specifically to recombinant human GM-CSF. The reliable and consistent performance of HEK-Blue™ GM-CSF cells makes them suitable for release assays of therapeutic molecules that inhibit GM-CSF signaling, such as Mavrilimumab , a monoclonal antibody targeting the α chain of GM-CSF receptor (see figures).

Key features

• Readily assessable STAT5-SEAP reporter activity
• Convenient readout using  QUANTI-Blue™ Solution
• High sensitivity to human (h) GM-CSFactivity
• Stability guaranteed for 20 passages

Applications

• Therapeutic development
• Drug screening
• Release assay

Granulocyte-macrophage colony-stimulating factor (GM-CSF, aka CSF2), although originally identified as a hematopoietic growth factor, is now regarded as a pleiotropic regulator of inflammation in the responses to pathogens, autoimmune diseases, and cancer [1, 2].

 More details

All InvivoGen products are for internal research use only, and not for human or veterinary use.

Figures

Validation of the expression of human GMRα by HEK-Blue™ GM-CSF cells.
5 x 10⁵ cells were incubated with either an APC-conjugated isotype control (blue) or an APC-conjugated Anti-GMRα mAb (red) for 1 hour. The binding affinity was then measured using flow cytometry.

Dose-response of HEK-Blue™ GM-CSF cells to recombinant GM-CSF. Cells were stimulated with increasing concentrations of recombinant human GM-CSF (hGM-CSF) and mouse GM-CSF (mGM-CSF). After overnight incubation, the STAT5 response was determined using QUANTI-Blue™ Solution, a SEAP detection reagent. The optical density (OD) at 650 nm is shown as mean ± SEM.

Dose-dependent inhibition of HEK-Blue™ GM-CSF cell response using Mavrilimumab biosimilar. Increasing concentrations of Anti-hGM-CSFR-hIgG4 (S228P)  (1 ng/ml - 2.5 µg/ml) were incubated with HEK‑Blue™ GM-CSF cells for 1 hour before the addition of recombinant human GM-CSF (100 pg/ml). After overnight incubation, SEAP activity in the cell culture supernatant was assessed using QUANTI-Blue™ Solution. Data are shown as the percentage of activity (mean ± SEM).

Response of HEK-Blue™ GM-CSF cells to a panel of cytokines. Cells were stimulated with various human and mouse recombinant cytokines: 0.1 ng/ml of hGM-CSF, 10 ng/ml of mGM-CSF, hIL‑6, hIL-27, or 1000 U/ml of hIFN-α. After overnight incubation, SEAP activity was assessed using QUANTI-Blue™ Solution. The optical density (OD) at 650 nm is shown as mean ± SEM.

Specifications

Cell type: Epithelial

Tissue origin: Human Embryonic Kidney

Target: GM-CSF

Specificity: Human

Reporter gene: SEAP

Antibiotic resistance: Blasticidin, Hygromycin, Zeocin®

Detection range: Human GM-CSF: 5 pg/ml - 100 ng/ml

Growth medium: Complete DMEM (see TDS)

Growth properties: Adherent

Mycoplasma-free: Verified using Plasmotest™

Quality control: Each lot is functionally tested and validated.

Additional information: The cell surface expression of human GMRα  (CD116) in this cell line has been validated using flow cytometry.

Contents

HEK-Blue™ GM-CSF Cells (hkb-hgmcsfr)

• 1 vial containing 3-7 x 10⁶ cells
• 1 ml Normocin™ (50 mg/ml)
• 2 x 1 ml of HEK-Blue Selection (250X)
• 1 ml of QB reagent and 1 ml of QB buffer (sufficient to prepare 100 ml of QUANTI-Blue™ Solution, a SEAP detection reagent).

HEK-Blue™ GM-CSF vial (hkb-hgmcsfr-av)

• 1 vial containing 3-7 x 10⁶ cells

Shipped on dry ice (Europe, USA, Canada and some areas in Asia)

Notification:  Reference #hkb-hgmcsfr-av can only be ordered together with reference #hkb-hgmcsfr.

Details

Cell line description

HEK-Blue™ GM-CSF cells were generated by stable transfection with the genes encoding for human GMRα (CD116), GMRβ (CD131), STAT5, as well as a STAT5-inducible secreted embryonic alkaline phosphatase (SEAP) reporter. The binding of GM-CSF to its receptor triggers a signaling cascade leading to the activation of STAT5 and the subsequent production of SEAP. This can be readily assessed in the supernatant using QUANTI-Blue™ Solution, a SEAP detection reagent.

HEK-Blue™ GM-CSF cells strongly respond to human, but not mouse, GM-CSF. Of note, these cells are not responsive to human IL-6, IL-27, and type I IFN (see figures).

GM-CSF background

The granulocyte-macrophage colony-stimulating factor (GM-CSF, aka CSF2) belongs to the β-common chain cytokine family [1]. Although originally identified as a hematopoietic growth factor, this cytokine is now regarded as a pleiotropic regulator of inflammation in the responses to pathogens, autoimmune diseases, and cancer [1, 2]. GM-CSF signalization requires a multimeric structure comprising four α chains (GMRα, aka CD116), four β chains (GMRβ aka CD131), and four cytokines. This 12-protein complex allows the juxtaposition of the intracellular Janus kinase 2 (JAK2) and activation of the signal transducer and activator of transcription 5 (STAT5) [1]. Other signaling pathways include ERK, NF-κB, and AKT pathways [2, 3]. 

The understanding of cellular and molecular mechanisms whereby GM-CSF exerts its varied functions is key for the development of therapeutic strategies (e.g. cancer vaccines, blocking antibodies) [1]. 

1. Dougan M. et al., 2019. GM-CSF, IL-3, and IL-5 family of cytokines: regulators of inflammation. Immunity. 50(4):796-811.
2. Zhan Y. et al., 2019. The Pleiotropic Effects of the GM-CSF Rheostat on Myeloid Cell Differentiation and Function: More Than a Numbers Game. Front Immunol. 102679.
3. Hamilton J.A., 2020. GM-CSF in inflammation. J. Exp . Med. 217(1):e20190945.

FAQ Cell Lines

Any questions about our cell lines ? Visit our frequently asked questions page