Committed to providing comprehensive support for global drug development

The interleukin-10 (IL-10)-related cytokine family includes several members such as IL-10, IL-19, IL-20, IL-22, IL-24, IL-26, IL-28A/B, and IL-29, which also belong to the 2α-spiral cytokine class. IL-19, IL-20, IL-22, IL-24, and IL-26 also belong to the IL-20 subfamily. The IL-20 subfamily is a core driver of skin inflammation (especially psoriasis) and also participates in the regulation of Th2 diseases (asthma, AD) and the tumor microenvironment.

Four heterodimer receptors serve as functional receptors for IL-20 family cytokines, and IL-24 signaling is transduced through two different heterodimer receptors. One type of IL-24 receptor is the type I IL-20 receptor (IL-20R), composed of IL-20RA and IL-20RB, which shares IL-19 and IL-20 as ligands. IL-24 and IL-20 can also transduce additional signals via the type II IL-20R, composed of IL-20RB and the IL-22 receptor subunit (IL-22RA1). The receptors for IL-22 and IL-26 are composed of IL-22RA1 and IL-20RA, respectively, and these two receptors share IL-10RB. Both type I and type II IL-20 receptors are expressed on non-immune cells, such as keratinocytes and bronchial epithelial cells, but not on immune cells. Therefore, it is believed that IL-24 cannot activate immune cells.

The HEK293 Cynomolgus IL20&IL22&IL24 Effector Reporter Cell model effectively simulates the signal transduction process of IL20/IL22/IL24 in vivo, as shown in the figure below.

Figure 1. Schematic diagram of the HEK293 Cynomolgus IL20&IL22&IL24 Effector Reporter Cell model.

Figure 2. Recombinant Cyno IL20&IL22&IL24 Triple Effector Reporter Cell constitutively expressing IL22RA.

Figure 3. Dose Response of Ligands in Cyno IL20&IL22&IL24 Triple Effector Reporter Cell(C30).


Figure 4. Inhibition of hIL20 Induced Reporter Activity by IL22Ra Blocking Ab or IL20 Neutralization Ab in Cyno IL20&IL22&IL24 Triple Effector Cell(C30).



Figure 5. Inhibition of Cyno IL22 Induced Reporter Activity by IL22Ra Blocking Ab or IL22 Neutralization Ab in Cyno IL20&IL22&IL24 Triple Effector Cell(C30).



Figure 6. Inhibition of hIL24 Induced Reporter Activity by IL22Ra Blocking Ab or IL24 Neutralization Ab in Cyno IL20&IL22&IL24 Triple Effector Cell (C30).

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 Cynomolgus IL20&IL22&IL24 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.