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 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, all of which belong to the class 2α helical cytokines. IL-19, IL-20, IL-22, IL-24, and IL-26 also belong to the IL-20 subfamily. All members of the IL-10 cytokine family signal through heterodimeric receptors composed of an α-chain and a β-chain.

IL-10 is a cytokine with anti-inflammatory properties that plays a central role in infection by limiting the immune response to pathogens, thereby preventing damage to the host. IL-10 is produced by a variety of cell types and exerts pleiotropic effects on them, including dendritic cells (DCs), macrophages, B cells, natural killer (NK) cells, Th1 and Th2 cells, Foxp3+ regulatory T (Treg) cells, and keratinocytes. IL-10 possesses anti-inflammatory and immunosuppressive effects, thereby playing a critical role in downregulating immune responses and preventing excessive inflammation. In particular, the function of monocytes and macrophages is extensively influenced by IL-10. IL-10 enhances the release of anti-inflammatory mediators by these cells and inhibits their ability to produce pro-inflammatory molecules. Furthermore, IL-10 suppresses the antigen-presenting capacity of monocytes, macrophages, and dendritic cells, while enhancing their tolerance-inducing, phagocytic, and clearance functions. IL-10 can also suppress Th1, Th2, and Th17-mediated immune responses by inhibiting the proliferation of CD4+ T cells and their ability to produce pro-inflammatory cytokines. Furthermore, it inhibits the secretion of pro-inflammatory mediators by neutrophils, eosinophils, and mast cells, as well as the development of mast cells. In contrast to these immunosuppressive properties, IL-10 has also been shown to stimulate the proliferation and cytotoxic activity of natural killer cells in the presence of IL-2, as well as the proliferation and lytic activity of CD8+ T cells in vitro, and to enhance B-cell proliferation, differentiation, and MHC class II expression.

 Excessive production of IL-10 has been shown to lead to immunodeficiency during infection or tumor growth, while a deficiency in IL-10 is associated with chronic inflammation and autoimmune diseases, including inflammatory bowel disease, psoriasis, and rheumatoid arthritis.

Figure 1. Schematic diagram of the IL-10 Effector Reporter Cell model

Figure 2. Recombinant IL10 Effector Reporter Cell constitutively expressing IL10RA and IL10RB.

Figure 3.Dose Response of Recombinant Human IL-10  Protein in IL10 Effector Reporter Cell(C1).

Figure 4.Inhibition of rhIL-10 Induced Reporter Activity by IL-10R Blocking Ab in IL 10 Effector Reporter Cell(C1).

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 IL10 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.