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GHSR1a, short for growth hormone secretagogue receptor type 1a, is a fully functional 7-transmembrane G protein-coupled receptor responsible for all known biological effects of ghrelin, including stimulating appetite, promoting growth hormone release, regulating energy balance, and glucose and lipid metabolism. GHSR1a uses the endogenous hormone ghrelin as its primary ligand. It couples to the Gq/11 protein, with the core signaling pathway being PLC-β, generating IP3 and mobilizing intracellular calcium ions, thereby triggering downstream cascade reactions. Furthermore, it can interact with pathways such as Gi/o or Gs in different cellular environments, forming a complex signal transduction network. This receptor is widely distributed in multiple tissues, including the hypothalamus, hippocampus, pancreas, and heart. Its signaling precisely regulates various key physiological functions such as growth hormone release, appetite, energy metabolism, and neuroprotection, making it an important drug target for the treatment of cachexia, obesity, and related metabolic and neurological diseases.



Figure 1. Schematic diagram of the GHSR1a HEK293 cell model.

Figure 2. Recombinant GHSR1a HEK293 stably expressing GHSR1a.

Figure 3. HTRF IP-One Assay with GHSR1a HEK293(C23).

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.