KC-3757

293T-CRE-Luc2-mouse-GLP1R Cell Line

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Background of 293T-CRE-Luc2-mouse-GLP1R Cell Line

GLP-1R (Glucagon-like peptide-1 receptor) is a G protein-coupled receptor that plays a key role in glucose homeostasis, by mediating the effects of the incretin hormone GLP-1 (glucagon-like peptide-1) on pancreatic beta cells, which results in insulin secretion. GLP-1R is also expressed in various other tissues including the brain, heart, lungs, and kidneys. It has been implicated in various physiological and pathophysiological processes such as energy metabolism, cardiovascular function, and inflammation.Several drugs that target GLP-1R have been developed for the treatment of type 2 diabetes, including GLP-1 receptor agonists such as exenatide, liraglutide, and dulaglutide, which stimulate GLP-1R signaling and enhance insulin secretion. GLP-1R antagonists have also been investigated for their potential use in the treatment of obesity and other metabolic disorders.

Specifications

Catalog Number	KC-3757
Cell Line Name	293T-CRE-Luc2-mouse-GLP1R Cell Line
Clone Number	1#
Host Cell Line	293T-CRE-Luc2
Description	Stable 293T-CRE-Luc2 cell line expressing exogenous mouse GLP1R 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+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-mouse-GLP1R Cell Line was generated using a lentiviral vector expressing GLP1R sequence.

Characterization

Figure 1. 293T-CRE-Luc2-GLP1R cell line were seed into the 96-well plate, and treated with GLP-1 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.Drucker DJ. Glucagon-like peptide-1 and the islet beta-cell: augmentation of cell proliferation and inhibition of apoptosis. Endocrinology. 2003;144(12):5145-5148. 2.Pyke C, Knudsen LB. The glucagon-like peptide-1 receptor--or not? Endocrinology. 2013;154(1):4-8. 3.Finan B, Ma T, Ottaway N, et al. Unimolecular dual incretins maximize metabolic benefits in rodents, monkeys, and humans. Sci Transl Med. 2013;5(209):209ra151.