KC-3769

293T-cyno-GPR84-Cell-Line

34744

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Background of 293T-cyno-GPR84-Cell-Line

GPR84 is an inflammation inducing receptor highly expressed on immune cells and is a member of the GPCR superfamily, expected to activate the activity of the urinary angiotensin II receptor. Expected to participate in the neuropeptide signaling pathway. It is a part of the receptor complex.

Specifications

Catalog Number	KC-3769
Cell Line Name	293T-cyno-GPR84-Cell-Line
Host Cell Line	293T
Description	Stable 293T clone expressing exogenous cyno-GPR84 gene
Quantity	One vial 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 1:4-1:8 every 2-3 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-cyno-GPR84-cell-line was generated using a lentiviral vector expressing the cyno-GPR84 sequence.

Characterization

Figure 1: Characterization of cyno-GPR84 overexpression in the 293T-cyno-GPR84 stable clone using qPCR.

Figure 2: Characterization of cyno-GPR84 in the 293T-cyno-GPR84 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/mL puromycin)in a 37°C water bath. 2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes. 3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol. 4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium. 5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet. 6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask. 7. Incubate the flask at 37°C, 5% CO₂ incubator. 8. Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 20% FBS + 10% DMSO) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium. 6. Aliquot 1 mL of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage.

References

1.Wei L, Tokizane K, Konishi H, Yu HR, Kiyama H. Agonists for G-protein-coupled receptor 84 (GPR84) alter cellular morphology and motility but do not induce pro-inflammatory responses in microglia. J Neuroinflammation. 2017 Oct 3;14(1):198. doi: 10.1186/s12974-017-0970-y. PMID: 28974234; PMCID: PMC5627487. 2. Aktar R, Rondinelli S, Peiris M. GPR84 in physiology-Many functions in many tissues. Br J Pharmacol. 2024 May;181(10):1524-1535. doi: 10.1111/bph.16206. Epub 2023 Aug 28. PMID: 37533166. 3. Forsman H, Dahlgren C, Mårtensson J, Björkman L, Sundqvist M. Function and regulation of GPR84 in human neutrophils. Br J Pharmacol. 2024 May;181(10):1536-1549. doi: 10.1111/bph.16066. Epub 2023 Mar 27. PMID: 36869866.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.