KC-1072

CHOK1-human-FCGR2B Cell Line

21483

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Background of CHOK1-human-FCGR2B Cell Line

FCGR2B, also known as CD32B, is an important inhibitory receptor of the Fc gamma receptor family, which plays vital roles in regulating various immune response processes and the pathogenesis of many infectious diseases. FCGR2B is highly expressed by B cells, basophils and at low levels on monocytes. FCGR2B decreased function is associated with systemic lupus erythematosus, rheumatoid arthritis, Goodpasture\'s disease, multiple sclerosis and others, that may be a target for monoclonal antibody therapy for autoimmune diseases as well as malignancies.

Specifications

Catalog Number	KC-1072
Cell Line Name	CHOK1-human-FCGR2B Cell Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human FCGR2B 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 + 10ug/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1 human FCGR2B cell line was generated using a lentiviral vector expressing the human FCGR2B sequence.

Characterization

Figure 1: Characterization of FCGR2B overexpression in the CHOK1 FCGR2B stable clone using FACS.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 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. Sun Z, Sun X, Yuan Y, Li H, Li X, Yao Z. FCGR2B as a prognostic and immune microenvironmental marker for gliomas based on transcriptomic analysis. Medicine (Baltimore). 2023 Sep 15;102(37):e35084. doi: 10.1097/MD.0000000000035084. PMID: 37713871; PMCID: PMC10508392.
2.Alee I, Chantawichitwong P, Leelahavanichkul A, Paludan SR, Pisitkun T, Pisitkun P. The STING inhibitor (ISD-017) reduces glomerulonephritis in 129.B6.Fcgr2b-deficient mice. Sci Rep. 2024 May 14;14(1):11020. doi: 10.1038/s41598-024-61597-z. PMID: 38745067; PMCID: PMC11094069.
3.Xu Z, Moreno-Giró À, Zhao D, Krämer A, Pandey RK, Xu B, Lundström SL, Holmdahl R. Fcgr2b and Fcgr3 are the major genetic factors for cartilage antibody-induced arthritis, overriding the effect of Hc encoding complement C5. Eur J Immunol. 2024 Apr;54(4):e2350659. doi: 10.1002/eji.202350659. Epub 2024 Feb 5. PMID: 38314895.