Unveil the Invisible-THP-1 Monocyte Model

THP-1 cells are a monocyte cell line derived from human peripheral blood, possessing morphological and functional characteristics (including cell differentiation markers) similar to those of primary human monocytes, and are widely used for research on inflammation and immune responses. In the field of glycobiology research, THP-1 cells can be used to mimic the inflammatory response and cell signaling pathways in a chronic high-glucose environment. It can be said that THP-1 cells are currently an ideal model for researching immunity and inflammation.

CD BioGlyco has constructed a powerful platform for the development of Glycobiology Disease Models and has accumulated deep experience in In Vitro Glycobiology Disease Model Screening. Here, we provide THP-1 monocyte-based in vitro screening services to our clients all over the world.

• We use culture plates in which THP-1 monocyte cells are cultured, to which inflammatory activators (e.g. lipopolysaccharide) are added to stimulate THP-1 cells and induce them to undergo differentiation.
  • Macrophage M1: The THP-1 can be differentiated into macrophages induced by phorbol ester (PMA) and can be reinduced by lipopolysaccharide (LPS) and IFN-γ to polarize M1, which expresses MD2, CD14, and MyD88 genes, and releases cytokines, such as TNF-α and IL-6, which is a typical model of inflammation.
  • Macrophage M2: Induction by IL-4, IL-13, and macrophages allows for M2 polarization and secretion of inhibitory cytokines such as TGF-β and IL-10, which is similar to the process of tissue repair and reconstruction in the later stages of inflammation.
  In the above procedure, we measure cytokine release from THP-1 cells in the cell-free supernatant obtained after centrifugation, and cytokine production is assessed by a specific enzyme-linked immunosorbent (ELISA) assay.
• Construction of a THP-1 knockout model
  We efficiently transfer Cas9 into THP-1 cells by nuclear transfection, and single clones are selected for culture after the completion of drug screening. Different clones are selected for target site amplification and sequencing validation respectively, and positive clones for gene knockdown are screened out. Reverse transcription-polymerase chain reaction (RT-PCR) is utilized to explore the key genes associated with pathogen clearance by macrophages.
  We also explore intracellular signaling pathways by establishing knockout and knock-in THP-1 cell models. We are knocking down MAVS, cGAS, and STING in THP-1 cells, respectively, and introducing dsDNA and RNA-DNA complexes into the cells, which can be used to explore the signaling pathways activated by RNA-DNA complexes in the immune response.

Fig.1 THP-1-based construction of cell models. (CD BioGlyco)

Publication Data

Frequently Asked Questions

• What is the THP-1 cell model?
  The THP-1 cell model is a human leukemic monocytic cell line isolated and established from the peripheral blood of patients with acute monocytic leukemia. THP-1 cells have morphological and functional characteristics similar to those of primary human monocytes, including markers of cell differentiation. It is widely used in research on monocyte- and macrophage-related mechanisms, signaling pathways, and nutrient and drug transport.
• How to test THP-1 cells for contamination?
  Methods for detecting whether THP-1 cells are contaminated typically include the following:
  • Visual observation: THP-1 cells should show growth in suspension, if abnormal wall adherence or change in morphology occurs, it may be a sign of contamination.
  • Biochemical testing: Use aseptic manipulation to detect the presence of bacterial growth in the culture medium. Inoculate the medium onto a solid medium and observe for bacterial colonization after incubation for some time.
  • Endogenous contamination detection: For fungal, yeast, or mold contamination, staining can be performed using specific stains (e.g., Giemsa stain) and then observed under a microscope.
  • PCR detection: Detection of specific microbial DNA using PCR technology, which is a more sensitive and specific method of detection.

Advantages

• We design the THP-1 monocyte model according to the nature and characteristics of glycobiology diseases.
• We have constructed different glycobiology disease models and are continuously engaged in innovative research and development to ensure efficient and high-quality in vitro screening services to our clients.
• Our glycobiology disease models undergo rigorous quality testing to ensure good stability.

CD BioGlyco has the technical expertise in the field of glycobiology disease model development to provide in vivo and in vitro model construction and screening services. Please feel free to contact us if you would like to inquire about the details of the THP-1 monocyte model in vitro screening service.