KC-6305

CHOK1-rat-IGF1R-low Cell Line

63961

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Background of CHOK1-rat-IGF1R-low Cell Line

IGF1R (Insulin Like Growth Factor 1 Receptor) is a Protein Coding gene. This receptor binds insulin-like growth factor with a high affinity. It has tyrosine kinase activity. The insulin-like growth factor I receptor plays a critical role in transformation events. Cleavage of the precursor generates alpha and beta subunits. It is highly overexpressed in most malignant tissues where it functions as an anti-apoptotic agent by enhancing cell survival. Alternatively spliced transcript variants encoding distinct isoforms have been found for this gene.

Specifications

Catalog Number	KC-6305
Cell Line Name	CHOK1-rat-IGF1R-low Cell Line
NCBI/UniProt Accession Number	NM_052807.3
Clone Number	11#
Host Cell Line	CHOK1
Description	CHOK1 cell line stably expressing exogenous rat-IGF1R 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% RPMI1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI1640 + 10% FBS + 10 μg/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

CHOK1-rat-IGF1R-low cell line was generated using a lentiviral vector expressing the rat-IGF1R sequence.

Characterization

Figure1: Characterization of rat-IGF1R overexpressing in CHOK1 stable clones using FACS.

Figure2: Characterization of CHOK1-rat-IGF1R-low cell line stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 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 9.0mL 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% 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

Mountzios G, Kostopoulos I, Kotoula V, Sfakianaki I, Fountzilas E, Markou K, Karasmanis I, Leva S, Angouridakis N, Vlachtsis K, Nikolaou A, Konstantinidis I, Fountzilas G. Insulin-like growth factor 1 receptor (IGF1R) expression and survival in operable squamous-cell laryngeal cancer. PLoS One. 2013;8(1):e54048. doi: 10.1371/journal.pone.0054048. Epub 2013 Jan 24. PMID: 23365645; PMCID: PMC3554755.
Dong Z, Yao M, Wang L, Yan X, Gu X, Shi Y, Yao N, Qiu L, Wu W, Yao D. Abnormal expression of insulin-like growth factor-I receptor in hepatoma tissue and its inhibition to promote apoptosis of tumor cells. Tumour Biol. 2013 Dec;34(6):3397-405. doi: 10.1007/s13277-013-0912-y. PMID: 23797814.
Fleuren ED, Versleijen-Jonkers YM, Heskamp S, Roeffen MH, Bouwman WH, Molkenboer-Kuenen JD, van Laarhoven HW, Oyen WJ, Boerman OC, van der Graaf WT. The strength of small: improved targeting of insulin-like growth factor-1 receptor (IGF-1R) with F(ab')₂-R1507 fragments in Ewing sarcomas. Eur J Cancer. 2013 Sep;49(13):2851-8. doi: 10.1016/j.ejca.2013.04.009. Epub 2013 May 8. PMID: 23664098.

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.