KC-2342

CHOK1-CD7-Cell-Line

23868

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Background of CHOK1-CD7-Cell-Line

CD7 encodes a transmembrane protein that is a member of the immunoglobulin superfamily. Expressed on the surface of all thymocytes, most mature T cells, and natural killer cells. It plays an important role in T cell interactions and T cell/B cell interactions during early lymphatic development.

Specifications

Catalog Number	KC-2342
Cell Line Name	CHOK1-CD7-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous CD7 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% RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640 + 10% FBS + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 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-CD7-cell-line was generated using a lentiviral vector expressing the CD7 sequence.

Characterization

Figure 1: Characterization of CD7 overexpression in the CHOK1-CD7 stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI 1640 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 1:6-1:8 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

1. Haftcheshmeh SM, Tajbakhsh A, Kazemi M, Esmaeili SA, Mardani F, Fazeli M, Sahebkar A. The clinical importance of CD4+ CD7- in human diseases. J Cell Physiol. 2019 Feb;234(2):1179-1189. doi: 10.1002/jcp.27099. Epub 2018 Aug 1. PMID: 30067877. 2.Sempowski GD, Lee DM, Kaufman RE, Haynes BF. Structure and function of the CD7 molecule. Crit Rev Immunol. 1999;19(4):331-48. PMID: 10530432.