KC-4050

293T FGFR3IIIC Cell Line

40558

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Background of 293T FGFR3IIIC Cell Line

Fibroblast growth factor receptor (FGFR) signalling plays an important role in embryogenesis as well as in tumorigenesis. In current studies FGFR has proved to be a potential molecular target in a variety of solid tumours.FGFR3 antibodies have been associated with sensory neuropathy, but many questions remain regarding their use in clinical practice.FGFR3 overexpression defines a subgroup of metastatic colorectal cancers with an unfavourable prognosis. Since FGFR3 alterations can present a potential therapeutic target, patients with FGFR3 overexpression should be included into clinical studies with FGFR inhibitors.

Specifications

Catalog Number	KC-4050
Cell Line Name	293T FGFR3IIIC Cell Line
Host Cell Line	293T
Description	Stable HEK293 cell line expressing exogenous human FGFR3IIIC 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1ug/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:5 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
In Vivo Validation	NA

Cell Line Generation

293T human FGFR3IIIC cell line was generated using a lentiviral vector expressing the FGFR3IIIC sequence.

Characterization

Figure 1: Characterization of human FGFR3IIIC overexpression in the 293T human FGFR3IIIC stable clone using FACS.

Figure2: Characterization of endogenous human FGFR3IIIC expression in 293T using FACS.

Cell Resuscitation

1. Prewarm culture medium (DMEM + 10% FBS + 1ug/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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 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.Latov N. FGFR3 autoantibodies in sensory neuronopathy. J Neurol Neurosurg Psychiatry. 2020 Jan;91(1):8. 2.Fromme JE, Schildhaus HU. FGFR3-Überexpression ist eine relevante Alteration in kolorektalen Karzinomen [FGFR3 overexpression is a relevant alteration in colorectal cancer]. Pathologe. 2018 Dec;39(Suppl 2):189-192. German.