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Chemokines belong to the cytokine family and possess chemotactic properties, regulating the migration and activation of phagocytes and lymphocytes. Chemokine receptors are G protein-coupled receptors (GPCRs) characterized by seven transmembrane domains that bind their ligands with high affinity. To date, at least 20 chemokine receptors have been identified, categorized into four subtypes: CXC chemokine receptors (CXCRs), CC chemokine receptors (CCRs), XCR, and CX3CR.

CCR7 (C-C motif chemokine receptor 7) is a G protein-coupled receptor with a seven-transmembrane structure, expressed primarily on the surface of T cells, B cells, and dendritic cells. Acting as a key "navigator" for the immune system, it recognizes and binds to the ligands CCL19 and CCL21, thereby activating intracellular signaling pathways such as PI3K/AKT. This process guides naive immune cells to home to lymph nodes and facilitates their interaction with antigen-presenting cells, ultimately initiating an adaptive immune response. However, this mechanism is frequently hijacked by various cancer cells, enabling them to migrate along the same signaling axis to lymph nodes and form metastases; consequently, CCR7 serves not only as a central molecule for maintaining immune homeostasis but also as a critical target in cancer therapy and autoimmune disease research.

The CHO-K1 Human CCR7 Cell Line model effectively simulates the in vivo CCR7 signal transduction process, the principle is illustrated in the figure below.

Figure 1. Schematic diagram of the CHO-K1 Human CCR7 Cell Line model

Figure 2. Recombinant CCR7/CHO-K1 constitutively expressing CCR7.

Figure 3. HTRF cAMP Assay with CCR7 CHO(C5).

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 CCR7 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.