KC-3443

293T-cyno-GIPR Cell Line

26509

Home » 293T-cyno-GIPR Cell Line

Background of 293T-cyno-GIPR Cell Line

GIPR, also named PGQTL2, is a G protein-coupled receptor that belongs to the 7-transmembrane protein family. GIPR is found on beta-cells in the pancreas,serving as the receptor for the Gastric inhibitory polypeptide hormone. Diseases associated with GIPR include Adrenal Cortical Adenoma and Adrenal Adenoma. Among its related pathways are GPCR downstream signaling and the Presynaptic function of Kainate receptor. Defects in this gene may contribute to the pathogenesis of diabetes.

Specifications

Catalog Number	KC-3443
Cell Line Name	293T-cyno-GIPR Cell Line
Clone Number	1-1#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous cyno GIPR 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 + 1ug/ml Puromycin
Selection Marker	Puromycin
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 cyno GIPR cell line was generated using lentiviral vector expressing cyno GIPR sequence.

Characterization

Figure 1: Characterization of cyno GIPR overexpression in the 293T cyno GIPR stable clone using FACS.

Cell Resuscitation

1. Prewarm culture medium (DMEM + 10% FBS + 1ug/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% 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.ntrez Gene: gastric inhibitory polypeptide receptor.Stoffel M, Fernald AA, Le Beau MM, Bell GI (August 1995). Assignment of the gastric inhibitory polypeptide receptor gene (GIPR) to chromosome bands 19q13.2-q13.3 by fluorescence in situ hybridization. Genomics. 28 (3): 607–609.. 2. Brubaker PL, Drucker DJ (2002). Structure-function of the glucagon receptor family of G protein-coupled receptors: the glucagon, GIP, GLP-1, and GLP-2 receptors.