KC-1068

CHOK1-FCGR3B-Cell-Line

21475

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Background of CHOK1-FCGR3B-Cell-Line

CD16, also named FC gamma RIII, is a low or intermediate affinity FC receptor, and has been identified as two receptors including FcγRIIIa (CD16a) and FcγRIIIb (CD16b). It is involved in phagocytosis, secretion of enzymes and inflammatory mediators, antibody-dependent cytotoxicity and clearance of immune complexes.

Specifications

Catalog Number	KC-1068
Cell Line Name	CHOK1-FCGR3B-Cell-Line
Host Cell Line	Chinese hamster ovary CHO-k1 cell line
Description	CHO-K1 cell line stable expressing exogenous human FCGR3B 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+5µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 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

CHO-K1 human FCGR3B cell line was generated using lentiviral vector expressing human FCGR3B sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (F12K + 10% FBS + 5µg/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 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% F12K + 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

Nimmerjahn F, Ravetch J (2006). "Fcgamma receptors: old friends and new family members". Immunity. 24 (1): 19ÿ28. doi:10.1016/j.immuni.2005.11.010. PMID 16413920.
Repp R, Valerius T, Sendler A, Gramatzki M, Iro H, Kalden J, Platzer E (1991). "Neutrophils express the high affinity receptor for IgG (Fc gamma RI, CD64) after in vivo application of recombinant human granulocyte colonystimulating factor". Blood. 78 (4): 885ÿ9. PMID 1714327
Ernst L, Duchemin A, Miller K, Anderson C (1998). "Molecular characterization of six variant Fcgamma receptor class I (CD64) transcripts". Mol Immunol. 35 (14ÿ15): 943ÿ54. PMID 9881691.