KC-4466

CT26-FGFR3IIIb Cell Line

46907

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Background of CT26-FGFR3IIIb Cell Line

FGFR3 is a tyrosine kinase receptor,Family members of the FGFR family are encoded by 4 different FGFR genes.The FGFR3 gene is located on the short arm of chromosome 4,It contains 19 exons and 18 introns with a span of 16.5kb.FGFR3 is usually mediated by gene amplification, activation of mutations, or bases Abnormal activation due to fusion or other methods.The FGFR3 gene is an arm One of the most commonly mutated genes in cystathione.

Specifications

Catalog Number	KC-4466
Cell Line Name	CT26-FGFR3IIIb Cell Line
Host Cell Line	CT26
Description	Stable CT26clone expressing exogenous FGFR3IIIb 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	Epithelial
Subculture	Split saturated culture 1:4~1:10 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	Yes

Cell Line Generation

CT26-FGFR3IIIb Cell Line was generated using lentivirus expressing cynomolgus FGFR3IIIb gene sequence.

Characterization

Figure 1: Characterization of FGFR3IIIb overexpression in the CT26-FGFR3IIIb stable clone using FACS.

Figure 2: Characterization of FGFR3IIIb overexpression in the CT26-FGFR3IIIb stable clone using PCR sequencing.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10ug/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 x 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 ~ 1:5 every 2~3 days; seed out at about 1-3 x 10
5 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 x g for 5 minutes at room temperature and carefully aspirate off the medium.
Resuspend the cells at a density of at least 3 x106 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

Tsai SJ, Wu MH, Chen HM, Chuang PC, Wing LY. Fibroblast growth factor-9 is an
endometrial stromal growth factor. Endocrinology. 2002 Jul;143(7):2715-21. doi:
1210/endo.143.7.8900. PMID: 12072406.
Sheeba CJ, Andrade RP, Duprez D, Palmeirim I. Comprehensive analysis of
fibroblast growth factor receptor expression patterns during chick forelimb
development. Int J Dev Biol. 2010;54(10):1517-26. doi: 10.1387/ijdb.092887cs.
PMID: 21302260.
Hafner C, Hartmann A. R248C-FGFR3-Mutation. Einfluss auf Zellwachstum,
Apoptose und Attachment in HaCaT-Keratinozyten [R248C FGFR3 mutation. Effect on
cell growth, apoptosis and attachment in HaCaT keratinocytes]. Pathologe. 2010
Oct;31 Suppl 2:221-4. German. doi: 10.1007/s00292-010-1332-z. PMID: 20711586.