KC-1402

CHOK1-CD24-Cell-Line

22069

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Background of CHOK1-CD24-Cell-Line

CD24, also known as heat stable antigen or small-cell lung carcinoma cluster 4 antigen, is a heavily glycosylated glycosylphosphatidylinositol-anchored surface protein and is expressed on B cells, T cells, keratinocytes, and myofiber synaptic nuclei and is upregulated in a wide variety of cancers. CD24 is a potent anti-phagocytic, "don’t eat me" signal that is capable of directly protecting cancer cells from attack by Siglec-10-expressing macrophages. CD24-Siglec10 as an innate immune checkpoint that is essential for mediating anti-tumor immunity and provides evidence for the therapeutic potential of CD24 blockade, with promise for the treatment of ovarian and breast cancers.

Specifications

Catalog Number	KC-1402
Cell Line Name	CHOK1-CD24-Cell-Line
Host Cell Line	Chinese hamster ovary CHO-K1 cell line
Description	CHOK1 cell line stable expressing exogenous human CD24 gene
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+10µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial-like
Subculture	Split saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL
Incubation	37 °C with 5% CO₂
Storage	Liquid nitrogen immediately upon receiving
Doubling Time	Approximately 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1 human CD24 cell line was generated using lentiviral vector expressing human CD24 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (F12K + 10% FBS + 10µg/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 1:4-1:8 every 2-3 days; seed out at about 1-2 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% F12K + 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

K Pirruccello, S. J. & LeBien, T. W. The human B cell-associated antigen CD24 is a single chain sialoglycoprotein. J. Immunol. 136, 3779ÿ3784 (1986).
Chen, G. Y., Brown, N. K., Zheng, P. & Liu, Y. Siglec-G/10 in selfÿnonself discrimination of innate and adaptive immunity. Glycobiology 24, 800ÿ806
Amira A. Barkal, et al. CD24 signaling through macrophage Siglec-10 is a target for cancer immunotherapy. Nature research.2019.