KC-1888

293T-GPRC5A-Cell-Line

22993

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Background of 293T-GPRC5A-Cell-Line

GPRC5A, also known as RAI3, is a protein that encodes a member of the type 3 G protein-coupled receptor family. GPRC5A may be involved in the interaction between retinoic acid and G protein signaling pathways. GPRC5A plays a critical role in development, cellular growth, and differentiation, and it may play a role in embryonic development and epithelial cell differentiation. GPRC5A is dysregulated in many human cancers and other diseases.

Specifications

Catalog Number	KC-1888
Cell Line Name	293T-GPRC5A-Cell-Line
Host Cell Line	Human HEK293T cell line
Description	Stable HEK293T cell line expressing exogenous human GPRC5A gene
Quantity	Two vials of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	70% DMEM+20% FBS+10% DMSO
Propagation Medium	DMEM+10%FBS+1µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 every 3 days; seed out at about 1-3 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

293T human GPRC5A cell line was generated using a lentiviral vector expressing the human GPRC5A sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (DMEM + 10% FBS + 1µg/mL Puromycin) in a 37°C water bath.
Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
Incubate the flask at 37°C, 5% CO₂ incubator.
Split saturated culture 1:3-1:6 every 3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use.
Keep the freezing medium on ice and label cryovials.
Transfer cells to a sterile, conical centrifuge tube, and count the cells.
Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
Aliquot 1 mL of the cell suspension into each cryovial.
Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
Transfer vials to liquid nitrogen for long-term storage

References

Cheng Y, Lotan R (1998). "Molecular cloning and characterization of a novel retinoic acid-inducible gene that encodes a putative G protein-coupled receptor". J. Biol. Chem. 273 (52): 35008ÿ15. doi:10.1074/jbc.273.52.35008. PMID 9857033.
"Entrez Gene: GPRC5A G protein-coupled receptor, family C, group 5, member A".
Cafferata EG, Gonzalez-Guerrico AM, Pivetta OH, Santa-Coloma TA (1996). "Identification by differential display of a mRNA specifically induced by 12-O-tetradecanoylphorbol-13-acetate (TPA) in T84 human colon carcinoma cells". Cell. Mol. Biol. (Noisy-le-grand). 42 (5): 797ÿ804. PMID 8832110.
Zhou H, Rigoutsos I (2014). "The emerging roles of GPRC5A in diseases". Oncoscience. 1 (12): 765ÿ76. doi:10.18632/oncoscience.104. PMC 4303886. PMID 25621293.