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	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	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.

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.