KC-1936

CHOK1-human-CD94-Cell-Line

23081

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Background of CHOK1-human-CD94-Cell-Line

CD94 is a protein-coding gene that is preferentially expressed on NK cells and is classified as a type II membrane protein due to its external C-terminus. CD94 plays an important role in immune response and self-tolerance mechanisms, and diseases associated with CD94 include Nk cell deficiency and chronic Nk cell lymphocytosis. The study of CD94 may provide new ideas for the diagnosis and treatment of these diseases.

Specifications

Catalog Number	KC-1936
Cell Line Name	CHOK1-human-CD94-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous human CD94 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
Subculture	Split saturated culture 1:4-1:5 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 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1 human CD94 Cell Line was generated using a lentiviral vector expressing the human CD94 sequence.

Characterization

Figure 1: Characterization of human CD94 overexpression in the CHOK1 human CD94 stable clone using FACS

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 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 1:4-1:5 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

Liu X, Song J, Zhang H, Liu X, Zuo F, Zhao Y, Zhao Y, Yin X, Guo X, Wu X, Zhang H, Xu J, Hu J, Jing J, Ma X, Shi H. Immune checkpoint HLA-E:CD94-NKG2A mediates evasion of circulating tumor cells from NK cell surveillance. Cancer Cell. 2023 Feb 13;41(2):272-287.e9. doi: 10.1016/j.ccell.2023.01.001. Epub 2023 Jan 26. PMID: 36706761.
Fang H, Wang W, Kadia TM, El Hussein S, Wang SA, Khoury JD. CD94 expression patterns in reactive and neoplastic T-cell and NK-cell proliferations. Leuk Res. 2021 Sep;108:106614. doi: 10.1016/j.leukres.2021.106614. Epub 2021 May 10. PMID: 33990003.