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IL-9 (interleukin-9) was initially identified as a T-cell growth factor and belongs to the IL-7/IL-9 cytokine family. Its receptor is a heterodimer composed of a specific IL-9 receptor alpha chain (IL-9Rα) and the common gamma chain (γc). IL-9 is produced by a wide range of cells, including adaptive immune cells (such as Th9, Th2, Th17, and Treg cells) and innate immune cells (such as mast cells, NKT cells, and ILC2s); among these, Th9 cells are recognized as the primary CD4+ T-helper cell subset responsible for IL-9 production.

Upon binding to its receptor, IL-9 primarily activates the JAK-STAT signaling pathway to regulate downstream gene expression, thereby influencing cell proliferation, differentiation, and survival. Functionally, IL-9 promotes the proliferation of T cells and mast cells and plays a pivotal role in driving allergic and inflammatory conditions (such as asthma) as well as autoimmune diseases (such as rheumatoid arthritis and systemic lupus erythematosus). Furthermore, IL-9 exerts a dual role in anti-tumor immunity, capable of both enhancing anti-tumor responses and potentially promoting tumor immune escape. These mechanisms position IL-9 as both a biomarker and a potential therapeutic target for various diseases, including asthma and COVID-19.

The HEK293 Human IL9 Effector Reporter Cell model effectively simulates the in vivo IL9 signal transduction process, the principle is illustrated in the figure below.

Figure 1. Schematic diagram of the HEK293 Human IL9 Effector Reporter Cell model

Figure 2. Recombinant hIL9 Effector Reporter Cell constitutively expressing IL9Rα&CD132.

Figure 3. Dose Response of Recombinant Human IL9 in IL9 Effector Reporter Cell(C18).

Figure 4. Inhibition of Human IL9-induced Reporter Activity by IL9 Neutralization Ab in IL9 Effector Reporter Cell (C18).

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 IL9 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.