KC-1614

Jurkat-CD47-KO-1B1-Cell-Line

22471

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Background of Jurkat-CD47-KO-1B1-Cell-Line

The therapeutic effect of CD47 blockade in syngeneic tumor models largely depends on the activation of T cells. More specifically, we demonstrate that the therapeutic effects of anti-CD47 relies on a cytosolic DNA sensor, dendritic cells (DCs), type I/II IFNs, and CD8 T cells. As such, we conclude that anti-CD47-mediated tumor rejection requires both innate and adaptive immune responses.

Specifications

Catalog Number	KC-1614
Cell Line Name	Jurkat-CD47-KO-1B1-Cell-Line
Host Cell Line	Jurkat
Description	Stable Jurkat clone with human CD47 gene knockout, No.1B1
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
Selection Marker	N/A
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-CD47-KO-1B1 cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of CD47 knockout in Jurkat using PCR sequencing.

Figure 2: Characterization of CD47 knockout in Jurkat using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS)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% 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

Liu X, Pu Y, Cron K, et al. CD47 blockade triggers T cell-mediated destruction of immunogenic tumors. Nat Med. 2015;21(10):1209-1215.
2.Boerma M, Freeman ML. Radiation Biology: Targeting CD47 in Cancer Growth Inhibition and Normal Tissue Protection. Int J Radiat Oncol Biol Phys. 2016;96(2):245-247.

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.