KC-1630

Jurkat-NFAT-Luc2-TIGIT-Cell-Line

22503

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Background of Jurkat-NFAT-Luc2-TIGIT-Cell-Line

T cell immunoglobulin and ITIM domain (TIGIT) is an inhibitory receptor expressed on lymphocytes that was recently propelled under the spotlight as a major emerging target in cancer immunotherapy. TIGIT interacts with CD155 expressed on antigen-presenting cells or tumour cells to down-regulate T cell and natural killer (NK) cell functions. Here we identify TIGIT as a coinhibitory receptor that critically limits antitumor and other CD8(+) T cell-dependent chronic immune responses. TIGIT is highly expressed on human and murine tumor-infiltrating T cells, and, in models of both cancer and chronic viral infection, antibody coblockade of TIGIT and PD-L1 synergistically and specifically enhanced CD8(+) T cell effector function, resulting in significant tumor and viral clearance, respectively.

Specifications

Catalog Number	KC-1630
Cell Line Name	Jurkat-NFAT-Luc2-TIGIT-Cell-Line
Host Cell Line	Jurkat
Description	Stable Jurkat cell line expressing exogenous luciferase gene under the control of TIGIT signaling pathway
Quantity	One vial of frozen cells (≥2-10⁶/vial)
Stability	Stable in culture over a minimum of 10 passages
Application	Cell model for monitoring EGFR signaling pathway
Freezing Medium	70% RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10%FBS+300µg/mL Hygromycin B+0.75µg/mL Puromycin
Selection Marker	Puromycin, HygromycinB
Morphology	Lymphoblast
Subculture	Split saturated culture 1:4-1:6 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-TIGIT cell line was generated using lentivirus expressing TIGIT sequence.

Characterization

Figure 1. Characterization of TIGIT over-expression in Jurkat-NFAT-Luc2 stable clones using FCAS.

Figure 2. Jurkat-NFAT-Luc2-TIGIT cell line was seeded into the 96-well plate, and co-culture with 293T-OS8-PDL1 for 6 hours, then readout with Bright-Glo system.

Figure 3.Jurkat-NFAT-Luc2-TIGIT cell line was seeded into the 96-well plate, and treated with KA-1177-Tiragolumab, KA-1016-Atezolizumab and KP2001-hIgG1 isotype at a maximum concentration 4 μg/mL for 1 hours, and co-culture with 293T-OS8-PDL1 for 6 hours, then readout with Bright-Glo system.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 300µg/mL Hygromycin B + 0.75µg/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 1:4-1:6 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

Harjunpää H, Guillerey C. TIGIT as an emerging immune checkpoint. Clin Exp Immunol. 2020 May;200(2):108-119.
Johnston RJ, Comps-Agrar L, Hackney J, Yu X, Huseni M, Yang Y, Park S, Javinal V, Chiu H, Irving B, Eaton DL, Grogan JL. The immunoreceptor TIGIT regulates antitumor and antiviral CD8(+) T cell effector function. Cancer Cell. 2014 Dec 8;26(6):923-937.

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.