KC-4448

CHOK1-mouse-GIPR Cell Line

46964

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Background of CHOK1-mouse-GIPR Cell Line

GIPR (Gastric Inhibitory Polypeptide Receptor) is a Protein Coding gene. This gene encodes a G-protein coupled receptor for gastric inhibitory polypeptide (GIP), which was originally identified as an activity in gut extracts that inhibited gastric acid secretion and gastrin release, but subsequently was demonstrated to stimulate insulin release in the presence of elevated glucose. Mice lacking this gene exhibit higher blood glucose levels with impaired initial insulin response after oral glucose load. Defect in this gene thus may contribute to the pathogenesis of diabetes.

Specifications

Catalog Number	KC-4448
Cell Line Name	CHOK1-mouse-GIPR Cell Line
Host Cell Line	CHOK1
Description	CHOK1 cell line stably expressing exogenous mouse 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10 μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Fibroblastoid cells growing as monolayer
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1-mouse-GIPR cell line was generated using a lentiviral vector expressing the mouse GIPR sequence.

Characterization

Figure1: Characterization of mouse GIPR overexpression in CHOK1 mouse GIPR stable clones using quantitative real-time PCR.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10 μg/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 9.0 mL 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 1-2 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Sauber J, Grothe J, Behm M, Scherag A, Grallert H, Illig T, Hinney A, Hebebrand J, Wiegand S, Grüters A, Krude H, Biebermann H. Association of variants in gastric inhibitory polypeptide receptor gene with impaired glucose homeostasis in obese children and adolescents from Berlin. Eur J Endocrinol. 2010 Aug;163(2):259-64. doi: 10.1530/EJE-10-0444. Epub 2010 Jun 1. PMID: 20516203.
Ismail S, Dubois-Vedrenne I, Laval M, Tikhonova IG, D\'Angelo R, Sanchez C, Clerc P, Gherardi MJ, Gigoux V, Magnan R, Fourmy D. Internalization and desensitization of the human glucose-dependent-insulinotropic receptor is affected by N-terminal acetylation of the agonist. Mol Cell Endocrinol. 2015 Oct 15;414:202-15. doi: 10.1016/j.mce.2015.07.001. Epub 2015 Jul 28. PMID: 26225752.