KC-1683

CHOK1-rat-FGFR2A-Cell-Line

22609

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Background of CHOK1-rat-FGFR2A-Cell-Line

FGFR2A, Also known as CD32a, it is a member of the immunoglobulin Fc receptor gene family found on the surface of many immune response cells. The protein encoded by this gene is a cell surface receptor found on phagocytic cells such as macrophages and neutrophils, involved in phagocytosis and clearance of immune complexes.

Specifications

Catalog Number	KC-1683
Cell Line Name	CHOK1-rat-FGFR2A-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous rat FGFR2A 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
In Vivo Validation	NA

Cell Line Generation

CHOK1-rat-FGFR2A-cell-line was generated using a lentiviral vector expressing the rat FGFR2A sequence.

Characterization

Figure 1: Characterization of rat FGFR2A overexpression in the CHOK1-rat-FGFR2A stable clone using FACS

Cell Resuscitation

Prewarm culture medium (RPMI 1640 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 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70%RPMI 1640 + 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

López-Martínez R, Albaiceta GM, Amado-Rodríguez L, Cuesta-Llavona E, Gómez J, García-Clemente M, Vázquez-Coto D, Alvarez V, Coto E. The FCGR2A rs1801274 polymorphism was associated with the risk of death among COVID-19 patients. Clin Immunol. 2022 Mar;236:108954. doi: 10.1016/j.clim.2022.108954. Epub 2022 Feb 9. PMID: 35149195; PMCID: PMC8824710.
Dahlqvist J, Fulco CP, Ray JP, Liechti T, de Boer CG, Lieb DJ, Eisenhaure TM, Engreitz JM, Roederer M, Hacohen N. Systematic identification of genomic elements that regulate FCGR2A expression and harbor variants linked with autoimmune disease. Hum Mol Genet. 2022 Jun 22;31(12):1946-1961. doi: 10.1093/hmg/ddab372. PMID: 34970970; PMCID: PMC9239749.