KC-4870

Jurkat-NFAT-Luc2-CD64a Cell Line

53440

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Background of Jurkat-NFAT-Luc2-CD64a Cell Line

This gene encodes a protein that plays an important role in the immune response. This protein is a high-affinity Fc-gamma receptor. The gene is one of three related gene family members located on chromosome 1. FCGR1A (Fc Gamma Receptor Ia) is a Protein Coding gene. Diseases associated with FCGR1A include Peritonitis and Pharyngitis. Among its related pathways are ADORA2B mediated anti-inflammatory cytokines production and Regulation of actin dynamics for phagocytic cup formation. Gene Ontology (GO) annotations related to this gene include obsolete signal transducer activity, downstream of receptor and IgG binding.

Specifications

Catalog Number	KC-4870
Cell Line Name	Jurkat-NFAT-Luc2-CD64a Cell Line
Host Cell Line	Jurkat-NFAT-Luc2
Description	Stable Jurkat cell line expressing exogenous luciferase under the control of NFAT responsive element and CD64a sequence.
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 +300μg/mL Hygromycin B + 1μg/mL puromycin
Selection Marker	Hygromycin B and Puromycin
Morphology	Lymphoblast
Subculture	Split saturated culture 1:4-1:5 every 2-3 days; seed out at about 1 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 30 hours
Mycoplasma Status	Negative

Cell Line Generation

Jurkat-NFAT-Luc2-CD64a Cell Line was generated using a lentiviral vector expressing the CD64a sequence.

Characterization

Figure 1: Jurkat-NFAT-Luc2-CD64a cells and SK-BR-3 cells were seeded into 96-well plates, treated with Herceptin(Cat# KB-1187, Kyinno) in different concentrations for 16 hours, and then read out using Bright-Glo Detection System.

Cell Resuscitation

Prewarm culture medium (RPMI 1640 supplemented with 10% FBS, 300µg/mL Hygromycin B and 1μg/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 1:4-1:5 every 2-3 days; seed out at about 1 × 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

1. Valerius T, Repp R, de Wit TP, Berthold S, Platzer E, Kalden JR, Gramatzki M, van de Winkel JG. Involvement of the high-affinity receptor for IgG (Fc gamma RI; CD64) in enhanced tumor cell cytotoxicity of neutrophils during granulocyte colony-stimulating factor therapy. Blood. 1993 Aug 1;82(3):931-9. PMID: 7687898. 2. Holzer K, Konietzny P, Wilhelm K, Encke A, Henrich D. Phagocytosis by emigrated, intra-abdominal neutrophils is depressed during human secondary peritonitis. Eur Surg Res. 2002 Jul-Aug;34(4):275-84. doi: 10.1159/000063071. PMID: 12145553.