KC-4024

Jurkat-NFAT-Luc2-CD2-KO-2A2-Cell-Line

40506

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Background of Jurkat-NFAT-Luc2-CD2-KO-2A2-Cell-Line

The glycoprotein CD2 is a costimulatory receptor expressed mainly on T and NK cells that binds to LFA3(CD58), a cell surface protein expressed on e.g., antigen-presenting cells. Its functions are two-fold: adhesion and activation. CD2 serves to facilitate conjugate formation between the T-lineage cell and its cognate partner via intermolecular interaction of CD2 and LFA-3 on the former and latter cells, respectively.CD2 has an important role in the formation and organization of the immunological synapse that is formed between T cells and antigen-presenting cells upon cell-cell conjugation and associated intracellular signaling. CD2 expression is upregulated on memory T cells as well as activated T cells and plays an important role in activation of memory T cells despite the coexistence of several other costimulatory pathways.

Specifications

Catalog Number	KC-4024
Cell Line Name	Jurkat-NFAT-Luc2-CD2-KO-2A2-Cell-Line
Host Cell Line	Jurkat-NFAT-Luc2-reporter
Description	Stable Jurkat-NFAT-Luc2-reporter clone with human CD2 gene knockout, No.2A2
Quantity	One vial of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Drug screening and biological assays
Freezing Medium	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS+300μg/mL Hygromycin B
Selection Marker	N/A
Morphology	lymphoblast
Subculture	Split saturated culture 1:5-1:8 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 26 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

Jurkat-NFAT-Luc2-CD2-KO-2A2 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of Jurkat-NFAT-Luc2-CD2-KO-2A2 cell line stable clone using PCR sequencing.

Figure 2: Characterization of Jurkat-NFAT-Luc2-CD2-KO-2A2 cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of Jurkat-NFAT-Luc2-CD2-KO-2A2 cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640+10% FBS+300μg/mL Hygromycin B)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 1:5-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% RPMI1640 + 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

Binder C, Cvetkovski F, Sellberg F, Berg S, Paternina Visbal H, Sachs DH, Berglund E, Berglund D. CD2 Immunobiology. Front Immunol. 2020 Jun 9;11:1090. doi: 10.3389/fimmu.2020.01090. PMID: 32582179; PMCID: PMC7295915.
Moingeon P, Chang HC, Sayre PH, Clayton LK, Alcover A, Gardner P, Reinherz EL. The structural biology of CD2. Immunol Rev. 1989 Oct;111:111-44. doi: 10.1111/j.1600-065x.1989.tb00544.x. PMID: 2576417.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.