KC-2149

CHOK1-FGFR2-Cell-Line

44931

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Background of CHOK1-FGFR2-Cell-Line

FGFR2, also known as CD332, is a member of the fibroblast growth factor receptor family, and the amino acid sequences between members of this family are highly conserved during evolution. FGFR family members differ from each other in ligand affinity and tissue distribution. A complete representative protein consists of an extracellular region composed of three immunoglobulin like domains, a single hydrophobic transmembrane segment, and a cytoplasmic tyrosine kinase domain. The extracellular portion of this protein interacts with fibroblast growth factor, triggering downstream signaling cascades that ultimately affect cell proliferation and differentiation. This specific family member is a high affinity receptor for acidic, alkaline, or keratinocyte growth factors, depending on the isotype.Mutations in this gene are associated with Crouzon syndrome, Pfeiffer syndrome, Craniosynostosis, Apert syndrome, Jackson-Weiss syndrome, Beare-Stevenson cutis gyrata syndrome, Saethre-Chotzen syndrome, and syndromic craniosynostosis.

Specifications

Catalog Number	KC-2149
Cell Line Name	CHOK1-FGFR2-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous FGFR2 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%RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640 + 10% FBS + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 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 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-FGFR2-cell-line was generated using a lentiviral vector expressing the FGFR2 sequence.

Characterization

Figure 1: Characterization of FGFR2 overexpression in the CHOK1-FGFR2 stable clone using FACS

Cell Resuscitation

Prewarm culture medium (RPMI 1640 supplemented with 10% FBS and 10μg/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 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70%RPMI 1640 + 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

Byron SA, Pollock PM. FGFR2 as a molecular target in endometrial cancer. Future Oncol. 2009 Feb;5(1):27-32. doi: 10.2217/14796694.5.1.27. PMID: 19243295.
Hibberd CE, Bowdin S, Arudchelvan Y, Forrest CR, Brakora KA, Marcucio RS, Gong SG. FGFR-associated craniosynostosis syndromes and gastrointestinal defects. Am J Med Genet A. 2016 Dec;170(12):3215-3221. doi: 10.1002/ajmg.a.37862. Epub 2016 Aug 2. PMID: 27481450; PMCID: PMC5503117.