KC-3765

CHOK1-CD66a-short-Cell-Line

34736

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Background of CHOK1-CD66a-short-Cell-Line

CD66a, also known as CEACAM1, is a transmembrane glycoprotein of the immunoglobulin superfamily and a key mediator in various physiological and pathological processes, including tumor diseases. It is expressed in various tissues, including epithelial cells, immune cells, and endothelial cells.

Specifications

Catalog Number	KC-3765
Cell Line Name	CHOK1-CD66a-short-Cell-Line
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous CD66a-short 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% 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 24 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-CD66a-short-cell-line was generated using a lentiviral vector expressing the CD66a-short sequence.

Characterization

Figure 1: Characterization of CD66a-short overexpression in the CHOK1-CD66a-short stable clone using FACS.

Figure 2: Characterization of CD66a-short overexpression in the CHOK1 stable clone using FACS.

Figure 3: Characterization of CD66a-short in the CHOK1-CD66a-short stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 supplemented with 10% FBS and 10μg/mL puromycin)in a 37°C water bath. 2. Thaw the frozen vial in a 37°C water bath for 1-2 minutes. 3. Transfer the vial into biosafety cabinet, and wipe the surface with 70% ethanol. 4. Unscrew the top of the vial and transfer the cell suspension gently into a sterile centrifuge tube containing 9.0mL complete culture medium. 5. Spin at ~ 125 × g for 5-7 minutes at room temperature, and discard the supernatant without disturbing the pellet. 6. Resuspend cell pellet with the appropriate volume of complete medium and transfer the cell suspension into a T25 culture flask. 7. Incubate the flask at 37°C, 5% CO₂ incubator. 8. Split saturated culture 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI 1640 + 20% FBS + 10% DMSO) fresh immediately before use. 2. Keep the freezing medium on ice and label cryovials. 3. Transfer cells to a sterile, conical centrifuge tube, and count the cells. 4. Centrifuge the cells at 250×g for 5 minutes at room temperature and carefully aspirate off the medium. 5. Resuspend the cells at a density of at least 3×10⁶ cells/mL in chilled freezing medium. 6. Aliquot 1 mL of the cell suspension into each cryovial. 7. Freeze cells in the CoolCell freezing container overnight in a -80°C freezer. 8. Transfer vials to liquid nitrogen for long-term storage.

References

1.Götz L, Rueckschloss U, Najjar SM, Ergün S, Kleefeldt F. Carcinoembryonic antigen-related cell adhesion molecule 1 in cancer: Blessing or curse? Eur J Clin Invest. 2024 Dec;54 Suppl 2(Suppl 2):e14337. doi: 10.1111/eci.14337. Epub 2024 Oct 25. PMID: 39451132; PMCID: PMC11646294. 2. Götz L, Rueckschloss U, Ergün S, Kleefeldt F. CEACAM1 in vascular homeostasis and inflammation. Eur J Clin Invest. 2024 Dec;54 Suppl 2(Suppl 2):e14345. doi: 10.1111/eci.14345. PMID: 39674877; PMCID: PMC11646292.

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.