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GPR88 is an orphan G protein-coupled receptor (GPCR) predominantly expressed in the striatum of the brain; consequently, it is also referred to as the "striatum-specific GPCR" (STRG). It is primarily localized on medium spiny neurons (MSNs) and co-localizes with dopamine D1 and D2 receptors. Although its endogenous ligand has yet to be identified, GPR88 is known to couple with the Gαi signaling pathway, thereby inhibiting the production of cAMP. It plays a pivotal role in regulating motor coordination, cognitive function, and emotional processing. Dysregulated expression of GPR88 has been implicated in various neuropsychiatric disorders—such as schizophrenia, Parkinson's disease, bipolar disorder, and drug addiction—and it is therefore considered a promising therapeutic target for central nervous system (CNS) diseases.

Figure 1. HTRF cAMP Assay with GPR88 HEK293(C16).

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 GPR88 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.