KC-3672

293T-CRE-Luc2-GLP2R Cell Line

34602

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Background of 293T-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-3672
Cell Line Name	293T-CRE-Luc2-GLP2R Cell Line
Clone Number	3#
Host Cell Line	293T-CRE-Luc2
Description	Stable 293T-CRE-Luc2 cell line expressing exogenous GLP2R 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+150µg/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

293T-CRE-Luc2-GLP2R Cell Line was generated using a lentiviral vector expressing GLP2R sequence.

Characterization

Figure 1. 293T-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 (DMEM supplemented with 10%FBS, 150µg/mL Hygromycin B 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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (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.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.

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.