CHO-K1 Human CXCR2 Gα15 Cell Line

For research use only. Not intended for human or animal clinical trials, therapeutic or diagnostic use.

Figure 1. Recombinant CXCR2 Gα15 CHO stably expressing CXCR2

Flow cytometry analysis was performed to detect CXCR2 expression. The blue curve represents CXCR2 Gα15 CHO (C23) stained with anti-CXCR2 antibody, while the red curve is the parental CHO control stained with the same antibody. The median fluorescence intensity of CXCR2-expressing cells reaches 3.80E5, indicating robust CXCR2 surface expression.

Figure 2. HTRF IP-One Assay with CXCR2 Gα15 CHO( C23)

Dose-response curve of human IL-8 measured by IP-One HTRF assay in CXCR2 Gα15 CHO (C23) stable cells. IL-8 triggers CXCR2-mediated signaling to reduce IP1 accumulation. The EC50 value is 18.66 ng/mL, demonstrating functional activity of the CXCR2 receptor in this cell line.
Figure 3. HTRF cAMP Assay with CXCR2 Gα15 CHO( C23)

Dose-response curve of human IL-8 determined by HTRF cAMP assay in CXCR2 Gα15 CHO (C23) stable cells. IL-8 activates CXCR2 and downstream Gα15 signaling, triggering cAMP accumulation. The EC50 value is 166.1 ng/mL, confirming the functional response of CXCR2 in this cell line.

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 CXCR2 Gα15 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.