KC-3674

CHOK1-CRE-Luc2-GLP2R-Cell-Line

34604

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Background of CHOK1-CRE-Luc2-GLP2R-Cell-Line

The peptide hormones of the glucagon-like peptide (GLP) family are playing an increasingly important role in the clinical aspects of human diseases. For example, GLP-1 receptor-targeted imaging of insulinomas and novel diabetes drugs based on GLP-1. Known nutritional and anti-inflammatory effects are also translated into the use of GLP-2 analogs to treat short bowel syndrome, Crohn’s disease and Inflammatory bowel disease. The GLP-2 receptor (GLP2R) mediates the actions of GLP-2, which belongs to the G protein-coupled receptor superfamily and is a member of the glucagon receptor family.

Specifications

Catalog Number	KC-3674
Cell Line Name	CHOK1-CRE-Luc2-GLP2R-Cell-Line
Clone Number	2#
Host Cell Line	CHOK1-CRE-Luc2
Description	Stable CHOK1-CRE-Luc2 cell line expressing exogenous GLP2R 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% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+10µg/mL Puromycin+750µg/ml Hygromycin B
Selection Marker	Puromycin \| Hygromycin B
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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

CHOK1-CRE-Luc2-GLP2R Cell Line was generated using a lentiviral vector expressing GLP2R sequence.

Characterization

Figure 1. CHOK1-CRE-Luc2-GLP2R cell line were seeded into the 96-well plate, and treated with GLP-2 for 6 hours, then readout with Bright-lite™ Luciferase Assay system.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS, 750μg/ml Hygromycin and 10µg/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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 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.Song B, Ge H, Pu C, Li N. GLP2-GLP2R signal affects the viability and EGFR-TKIs sensitivity of PC9 and HCC827 cells. BMC Pulm Med. 2022 Jan 13;22(1):36. doi: 10.1186/s12890-021-01800-3. PMID: 35027025; PMCID: PMC8756716.