KC-1613

Jurkat-CD47-KO-1A3-Cell-Line

22469

Home » Jurkat-CD47-KO-1A3-Cell-Line

Background of Jurkat-CD47-KO-1A3-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-1613
Cell Line Name	Jurkat-CD47-KO-1A3-Cell-Line
Host Cell Line	Jurkat
Description	Stable Jurkat clone with human CD47 gene knockout, No.1A3
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	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-1A3 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.