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CCR2 (C-C chemokine receptor type 2) is a G protein-coupled receptor (GPCR) expressed on the surface of immune cells such as monocytes and macrophages. It features a seven-transmembrane helix structure, with a glycosylation site on the extracellular N-terminus and a protein interaction domain responsible for signal transduction on the intracellular C-terminus. The human CCR2 protein consists of 374 amino acids and has two primary splice variants—CCR2A and CCR2B—with CCR2B being the most common functional form.

The primary endogenous ligand for CCR2 is CCL2 (MCP-1). Upon binding to CCR2, CCL2 activates the Gαi signaling pathway, triggering downstream cascades involving β-arrestin and MAPK, which drive the migration of immune cells to sites of inflammation or injury. Under pathological conditions, overactivation of the CCR2/CCL2 axis promotes monocyte infiltration into tissues (such as in fatty liver, atherosclerotic plaques, and the tumor microenvironment) and mediates chronic inflammation and fibrosis; consequently, it has become a prominent therapeutic target for metabolic diseases (such as NASH), autoimmune disorders, pain, and cancer metastasis.

The CHO-K1 Human CCR2 β-Arrestin Cell Line model effectively simulates the in vivo CCR2 β-Arrestin signal transduction process, the principle is illustrated in the figure below.

Figure 1. Schematic diagram of the CHO-K1 Human CCR2 β-Arrestin Cell Line model

Figure 2. Recombinant CCR2/β-Arrestin/CHO-K1 constitutively expressing CCR2.

Figure 3. Dose response of Human CCL2 in CCR2 β-Arrestin CHO-K1(C10).

Figure 4. Inhibition of hCCL2-induced β-Arrestin Recruitment By CCL2 Neutralization Ab in CCR2 β-Arrestin CHO-K1(C10). Inhibition of hCCL2-induced β-Arrestin Recruitment in CCR2 β-Arrestin CHO-K1(C10).

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 CCR2 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.