KC-6372

293T-FCGR2A Cell Line

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Background of 293T-FCGR2A Cell Line

FCGR2A (Fc Gamma Receptor IIa) is a Protein Coding gene，it encodes one member of a family of immunoglobulin Fc receptor genes 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, and is involved in the process of phagocytosis and clearing of immune complexes. Diseases associated with FCGR2A include Cystic Fibrosis and Systemic Lupus Erythematosus.

Specifications

Catalog Number	KC-6372
Cell Line Name	293T-FCGR2A Cell Line
NCBI/UniProt Accession Number	P12318
Clone Number	5#
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous human FCGR2A 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 +1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:5 every 2-3 days
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-FCGR2A cell line was generated using a lentiviral vector expressing the human FCGR2A sequence.

Characterization

Figure 1: Characterization of human FCGR2A overexpression in the 293T-FCGR2A stable clone using FACS.

Figure 2: Characterization of human FCGR2A in the 293T-FCGR2A stable clone using PCR sequencing.

Cell Resuscitation

Pre-warm complete culture medium (DMEM+10% FBS +1μg/mL Puromycin) in a 37°C water bath.
Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
Subculture the cells at a ratio of 1:4-1:5 every 2-3 days upon reaching 80%–90% confluency.

Cell Freezing

Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
Pre-chill the freezing medium on ice and label the cryovials accordingly.
Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
Gently resuspend the cell pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

1. Li S, Shen W, Yu T, Ran F, Song Z, Yao Q, Zha Y. FCGR2A contributes to M2 macrophages polarization in HCC through IL-4/JAK/STAT6 axis. Transl Oncol. 2025 Aug;58:102429. doi: 10.1016/j.tranon.2025.102429. Epub 2025 Jun 6. PMID: 40482466; PMCID: PMC12177172.
2. Li S, Shen W, Yu T, Ran F, Song Z, Yao Q, Zha Y. FCGR2A contributes to M2 macrophages polarization in HCC through IL-4/JAK/STAT6 axis. Transl Oncol. 2025 Aug;58:102429. doi: 10.1016/j.tranon.2025.102429. Epub 2025 Jun 6. PMID: 40482466; PMCID: PMC12177172.
3. Fu T, Zhang L, Zuo M, Li F, Shi C, Chen H. FCGR2A as one novel potential target for poor survival prognosis of clear cell renal cell carcinoma. Medicine (Baltimore). 2023 Mar 17;102(11):e33324. doi: 10.1097/MD.0000000000033324. PMID: 36930102; PMCID: PMC10019103.