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Chemokine Receptor 9 (CCR9) belongs to the β-chemokine receptor family. It consists of two transcripts: Transcript A (369 amino acids, 42 kDa) and Transcript B (357 amino acids, 40.8 kDa). CCR9 is predominantly expressed on the surface of immature T lymphocytes and enterocytes; upon binding to its specific ligand, it plays a role in T-lymphocyte development and tissue-specific homing. Studies have demonstrated that CCR9 is highly expressed in various types of cancer, serving as a potential tumor biomarker applicable to both diagnosis and therapy.

Upon activation by its specific ligand, CCL25, CCR9 can interact with numerous signaling pathways—particularly those implicated in tumor chemoresistance and metastasis.

The CHO-K1 Mouse CCR9 β-Arrestin Cell Line model effectively mimics the in vivo CCR9 signaling transduction process. The principle is illustrated in the figure below.

Figure 1.  Schematic diagram of the CHO-K1 Mouse CCR9 β-Arrestin Cell Line model

Figure 2. Recombinant Mouse CCR9 β-Arrestin CHO stably expressing CCR9.

Figure 3. Dose Response of Mouse CCL25 in Mouse CCR9 β-Arrestin CHO(C89）.

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 Mouse CCR9 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.