Committed to providing comprehensive support for global drug development

The *IL10RA* gene (encoding Interleukin-10 receptor subunit alpha, also known as CD210 or IL-10R1) is located at the 11q23.3 region on the long arm of human chromosome 11. It encodes a transmembrane protein composed of 578 amino acids that is widely expressed in over a dozen tissues, including the spleen (RPKM 29.9) and bone marrow (RPKM 20.8). As the ligand-binding subunit of the IL-10 receptor complex, it must form a heterotetramer with the accessory subunit IL-10RB to facilitate signal transduction. Its primary function is to mediate the immunosuppressive signaling of IL-10: upon binding to IL-10, IL-10RA activates JAK1 and TYK2 kinases, which subsequently phosphorylate STAT3, inducing its dimerization and translocation into the nucleus; this process inhibits the expression of pro-inflammatory factors such as TNF-α, IL-6, and IL-1β, while simultaneously promoting the activation of regulatory T cells to maintain intestinal immune homeostasis.

Figure 1. Recombinant IL10RA  CHO stably expressing IL10RA .

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  CHO-K1 Human IL10RA 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.