KC-4095

CHOK1-CD2-Cell-Line

41799

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Background of CHOK1-CD2-Cell-Line

CD2 encodes a bifunctional protein. In the cytoplasm, the encoded protein binds the cytoplasmic tail of the human surface antigen CD2 through its C-terminal GYF structural domain and regulates CD2-triggered T lymphocyte activation. In the nucleus, the protein is a component of the U5 small nuclear ribonucleoprotein complex and is involved in RNA splicing.

Specifications

Catalog Number	KC-4095
Cell Line Name	CHOK1-CD2-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous CD2 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 1:4-1:8 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 CD2 Cell Line was generated using a lentiviral vector expressing the CD2 sequence.

Characterization

Figure 1: Characterization of CD2 overexpression in the CHO-K1 CD2 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 1:4-1:8 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

Binder C, Cvetkovski F, Sellberg F, Berg S, Paternina Visbal H, Sachs DH, Berglund E, Berglund D. CD2 Immunobiology. Front Immunol. 2020 Jun 9;11:1090. doi: 10.3389/fimmu.2020.01090. PMID: 32582179; PMCID: PMC7295915.
Ghosh A, Marques-Piubelli ML, Wang X, Sheu TG, Cheng J, Khan K, Lu W, Manning J, Tang G, Solis LM, Vega F. CD2-negative lymphoma-associated T-cells: a potential mechanism of immune-evasion in diffuse large B-cell lymphoma. Virchows Arch. 2022 Oct;481(4):659-663. doi: 10.1007/s00428-022-03348-x. Epub 2022 May 27. PMID: 35622145.
Kanatsu-Shinohara M, Chen G, Morimoto H, Shinohara T. CD2 is a surface marker for mouse and rat spermatogonial stem cells. J Reprod Dev. 2020 Aug 20;66(4):341-349. doi: 10.1262/jrd.2020-019. Epub 2020 Mar 26. PMID: 32213736; PMCID: PMC7470899.