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Interleukin-23 (IL-23) is a key pro-inflammatory cytokine involved in the pathogenesis of chronic inflammatory diseases—such as psoriasis, inflammatory bowel disease, multiple sclerosis, and rheumatoid arthritis—and belongs to the IL-12 family, alongside IL-12, IL-27, IL-35, and IL-39. The pathological consequences of excessive IL-23 signaling are linked to its capacity to induce target cell populations—primarily TCRγδ cells that secrete Th17 and IL-17 (Tγδ17 cells)—to produce inflammatory mediators, such as IL-17, IL-22, granulocyte-macrophage colony-stimulating factor (GM-CSF), or tumor necrosis factor (TNF-α). These pathogenic mediators promote the recruitment and activation of granulocytes and macrophages, leading to tissue damage, the induction of chronic inflammation, and ultimately, the manifestation of clinical symptoms.

IL-23 is a heterodimeric cytokine composed of two disulfide-linked subunits: p40 (encoded by *IL12B*) and p19 (*IL23A*). These subunits bind to a membrane receptor complex formed by IL-12Rβ1 and the unique IL-23R. IL-23 shares the p40 subunit, as well as one of its receptor components—IL-12Rβ1—with IL-12. Despite sharing a common subunit, IL-23 and IL-12 exert distinct biological functions. IL-23 stimulates the activity of JAK2 and TYK2, thereby leading to the activation of Signal Transducers and Activators of Transcription (STATs).

The HEK293 Human IL23 Effector Reporter Cell Model—effectively simulates the signal transduction process of  IL23 *in vivo*. The underlying principle is illustrated in the figure below.

Figure 1. Schematic Diagram of the HEK293 Human IL23 Effector Reporter Cell Model

HEK293 cell line expressing full length human IL23. Expression is confirmed by flow cytometry.

Figure 2. Recombinant IL23 Effector Reporter Cell stably expressing IL23R& IL12β1.

Figure 3.Dose Response of Recombinant Human IL23 in IL23 Effector Reporter Cell(C6).

Figure 4.Inhibition of rhIL23-induced Reporter Activity by IL23 Neutralizing Antibody in IL23 Effector Reporter Cell(C6).

Cell Resuscitation
1)Rapidly thaw the frozen cells in a 37 °C water bath for approximately 60 seconds. Once thawed (which may take slightly less or more than 60 seconds), immediately transfer the cell suspension from the cryovial into a 15 mL centrifuge tube containing 10 mL of pre-warmed  HEK293 Human IL23 Effector Reporter Cell complete culture medium.
2)Centrifuge cells at 1000 rpm for 5 min to remove medium, then resuspend cells in 5 mL of pre-warmed complete medium.
3)Transfer the cell suspension into a T25 culture flask and incubate at 37 °C with 5% CO₂.
4)After approximately 24–36 hours, replace the medium or passage the cells to remove non-adherent dead cells.

Subculturing procedure
1)When the cell density reaches the appropriate confluency for passaging, wash the cells with PBS, then add 1 mL trypsin to detach the cells. When more than 80% of the cells detach upon gently tapping the culture flask, add complete culture medium to terminate digestion. Gently pipette to obtain a single-cell suspension, transfer to a 15 mL centrifuge tube, and centrifuge at 1000 rpm for 5 minutes.

2)Discard supernatant after centrifugation. Resuspend cells in fresh medium to a single-cell suspension and transfer to a new culture flask for continued growth.

Cell Freezing
After trypsinization and centrifugation of cells from each T75 flask or 10 cm culture dish, discard the supernatant. Add 2 mL of cryopreservation medium (90% FBS + 10% DMSO), gently resuspend thoroughly, and aliquot into two cryovials. Immediately place the cryovials into a controlled-rate freezing container (e.g., Nalgene 5100-0001), fill with isopropanol to the indicated level, and store at −80 °C. After 24 hours, transfer the cryovials to liquid nitrogen for long-term storage.