KC-6461

293T-rat-GIPR Cell Line

62891

Home » 293T-rat-GIPR Cell Line

Background of 293T-rat-GIPR Cell Line

Human ortholog(s) of GIPR implicated in cardiovascular system disease; diabetes mellitus; and obesity. Orthologous to human GIPR (gastric inhibitory polypeptide receptor). Enables gastric inhibitory peptide receptor activity and peptide hormone binding activity. Involved in several processes, including desensitization of G protein-coupled receptor signaling pathway; gastric inhibitory peptide signaling pathway; and positive regulation of insulin secretion.

Specifications

Catalog Number	KC-6461
Cell Line Name	293T-rat-GIPR Cell Line
NCBI/UniProt Accession Number	XM_063281146.1
Clone Number	1#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous rat-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 +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

Cell Line Generation

293T-rat-GIPR cell line was generated using lentivirus expressing rat-GIPR sequence.

Characterization

Figure 1. Characterization of rat-GIPR over-expression in the 293T-rat-GIPR stable clone using FACS.

Figure 2. Characterization of 293T-rat-GIPR cell line stable clone using PCR sequencing.

Cell Resuscitation

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

Cell Freezing

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

References

Zhou J, Livak MF, Bernier M, Muller DC, Carlson OD, Elahi D, Maudsley S, Egan JM. Ubiquitination is involved in glucose-mediated downregulation of GIP receptors in islets. Am J Physiol Endocrinol Metab. 2007 Aug;293(2):E538-47. doi: 10.1152/ajpendo.00070.2007. Epub 2007 May 15. PMID: 17505054; PMCID: PMC2640485.