HEK293 Human GHSR1a Cell Line

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

Figure 1. Recombinant GHSR1a HEK293 stably expressing GHSR1a

The GHSR1a/HEK293 stable cells (blue histogram) showed a marked right shift in FL4-A fluorescence (median = 8897) compared with parental HEK293 cells (red histogram, median = 453), confirming robust and stable membrane expression of human GHSR1a.

Figure 2. HTRF IP-One Assay with GHSR1a HEK293(C23)

The GHSR1a stable HEK293 (C23, red squares) exhibited potent agonist response with an EC₅₀ of 0.60 nM. Parental HEK293 cells (blue circles) showed no detectable ghrelin-induced IP1 accumulation, with EC₅₀ > 2000 nM. The results confirm specific GHSR1a Gq signaling activation upon ghrelin stimulation.

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 HEK293 Human GHSR1a 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.