KC-5136

CHOK1-FGFR1b-Cell-Line

57809

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Background of CHOK1-FGFR1b-Cell-Line

FGFR1b is an important splicing variant of the FGFR1 gene, primarily expressed in epithelial cells, specifically binding to ligands such as FGF10 and playing a critical role in organ development and tissue homeostasis. Its abnormal activation is one of the driving factors for various cancers, especially epithelial derived cancers, and therefore has become an important research object for targeted cancer therapy. Understanding FGFR1b and its differences from FGFR1c is of great significance for precision medicine and drug development.

Specifications

Catalog Number	KC-5136
Cell Line Name	CHOK1-FGFR1b-Cell-Line
Clone Number	8-15#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous FGFR1b 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

Cell Line Generation

CHOK1-FGFR1b-cell-line was generated using a lentiviral vector expressing the FGFR1b sequence.

Characterization

Figure 1: Characterization of FGFR1b overexpression in the CHOK1-FGFR1b stable clone using FACS.

Figure 2: Characterization of FGFR1b in the CHOK1-FGFR1b stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 supplemented with 10% FBS and 10μg/mL puromycin)in a 37°C water bath.
2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes.
3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol.
4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium.
5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet.
6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask.
7. Incubate the flask at 37°C, 5% CO₂ incubator.
8. Split saturated culture 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI 1640 + 20% FBS + 10% DMSO) fresh immediately before use.
2. Keep the freezing medium on ice and label cryovials.
3. Transfer cells to a sterile, conical centrifuge tube, and count the cells.
4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium.
6. Aliquot 1 mL of the cell suspension into each cryovial.
7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer.
8. Transfer vials to liquid nitrogen for long-term storage.

References

1.Lee JM, Kim JY, Cho KW, Lee MJ, Cho SW, Kwak S, Cai J, Jung HS. Wnt11/Fgfr1b cross-talk modulates the fate of cells in palate development. Dev Biol. 2008 Feb 15;314(2):341-50. doi: 10.1016/j.ydbio.2007.11.033. Epub 2007 Dec 7. PMID: 18191119.
2.Dabrowski A, Terauchi A, Strong C, Umemori H. Distinct sets of FGF receptors sculpt excitatory and inhibitory synaptogenesis. Development. 2015 May 15;142(10):1818-30. doi: 10.1242/dev.115568. Epub 2015 Apr 29. Erratum in: Development. 2024 Aug 15;151(16):dev203123. doi: 10.1242/dev.203123. PMID: 25926357; PMCID: PMC4440923.