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Chemokine (C-C motif) receptor 8 (CCR8) is a G protein-coupled receptor primarily expressed on regulatory T cells (Tregs). Its ligands include CCL1, CCL8, CCL16, and CCL18.

CCR8 is specifically expressed on tumor-infiltrating Tregs, as well as on Th2 cells and tumor cells; however, it is largely absent from peripheral blood Tregs and normal tissues. Its primary function involves the recruitment of Tregs and Th2 cells to sites of inflammation and tumors. Numerous studies have demonstrated that high CCR8 expression is associated with various cancers—including colorectal, breast, gastric, metastatic brain, and metastatic liver cancers—with particularly high levels observed on Tregs isolated from breast cancer. Consequently, CCR8 has emerged as a significant research target that has garnered considerable attention in recent years.

The CHO-K1 Human CCR8 β-Arrestin Cell Line model effectively simulates the in vivo CCR8 β-Arrestin signal transduction process, the principle is illustrated in the figure below.

Figure 1. Schematic diagram of the CHO-K1 Human CCR8 β-Arrestin Cell Line model

Figure 1. Recombinant CCR8 β-Arrestin/CHO-K1 stably expressing CCR8.

Figure 2. Dose response of Human Recombinant CCL1 in CCR8 β-Arrestin CHO-K1(C6).

Figure 3. Inhibition of  rhCCL1-induced β-Arrestin Recruitment in CCR8 β-Arrestin CHO-K1(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 CHO-K1 Human CCR8 β-Arrestin Cell Line 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.