KC-2140-DW

CT26-FGFR2B Cell Line

57889

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Background of CT26-FGFR2B Cell Line

FGFR2B is a specific isoform of the Fibroblast Growth Factor Receptor 2, generated by alternative splicing of the FGFR2 gene. It is characterized by its expression primarily in epithelial cells and its unique ligand-binding specificity, primarily interacting with FGF ligands from the FGF7 subfamily (e.g., FGF7, FGF10, FGF22). The activation of FGFR2B by its ligands triggers crucial downstream signaling pathways (e.g., MAPK, PI3K-AKT) that regulate key cellular processes including proliferation, differentiation, migration, and survival during development and tissue homeostasis. Dysregulation of FGFR2B signaling—through gene amplification, overexpression, or mutations—has been implicated in the pathogenesis of various cancers, such as gastric, breast, and endometrial cancer, making it a potential therapeutic target for selective inhibitors.

Specifications

Catalog Number	KC-2140-DW
Cell Line Name	CT26-FGFR2B Cell Line
Host Cell Line	CT26
Description	Stable CT26 cell line expressing exogenous FGFR2B 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 + 10μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Fibroblastoid
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

CT26-FGFR2B Cell Line was generated using a lentiviral vector expressing the FGFR2B sequence.

Characterization

Figure 1: Characterization of CT26 over expression in the CT26-FGFR2B stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 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 1:4-1:8 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

Tartaglia M, Valeri S, Velardi F, Di Rocco C, Battaglia PA. Trp290Cys mutation in exon IIIa of the fibroblast growth factor receptor 2 (FGFR2) gene is associated with Pfeiffer syndrome. Hum Genet. 1997 May;99(5):602-6. doi: 10.1007/s004390050413. PMID: 9150725.
Mundhofir FE, Sistermans EA, Faradz SM, Hamel BC. p.Ser252Trp and p.Pro253Arg mutations in FGFR2 gene causing Apert syndrome: the first clinical and molecular report of Indonesian patients. Singapore Med J. 2013 Mar;54(3):e72-5. doi: 10.11622/smedj.2013055. PMID: 23546041.