KC-4270

A549-FGFR1-KO-1B1-Cell-Line

43818

Home » A549-FGFR1-KO-1B1-Cell-Line

Background of A549-FGFR1-KO-1B1-Cell-Line

FGFR1 (Fibroblast Growth Factor Receptor 1) is a member of the fibroblast growth factor receptor family, which is involved in various biological processes, such as cell growth, differentiation, and tissue repair. It is a receptor tyrosine kinase that binds to fibroblast growth factors (FGFs) and triggers a cascade of downstream signaling pathways including MAPK/ERK, PI3K/AKT, and PLCγ, promoting cellular responses. FGFR1 plays an essential role in embryonic development, angiogenesis, and neurogenesis. Dysregulation of FGFR1 has been implicated in various cancers, developmental disorders, and other diseases, making it a target for therapeutic interventions.

Specifications

Catalog Number	KC-4270
Cell Line Name	A549-FGFR1-KO-1B1-Cell-Line
Host Cell Line	A549
Description	Stable A549 clone with human FGFR1 gene knockout, No.1B1
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	McCoy5A+20% FBS+10% DMSO
Propagation Medium	McCoy5A+10% FBS
Selection Marker	N/A
Morphology	Epithelial
Subculture	Split saturated culture 1:3-1:6 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

A549-FGFR1-KO-1B1 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of A549-FGFR1-KO-1B1 cell line stable clone using PCR sequencing.

Figure 2: Characterization of A549-FGFR1-KO-1B1 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of A549-FGFR1-KO-1B1 cell line stable clone using western blot.

Cell Resuscitation

Prewarm culture medium (McCoy5A + 10% FBS)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 1:3-1:6 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% McCoy5A + 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

Turner, N., & Grose, R. (2010). Fibroblast growth factor signalling: from development to cancer. Nature Reviews Cancer, 10(2), 116–129. https://doi.org/10.1038/nrc2780