KC-3251

CT26-luc2-CEA6-CEA5-Cell-Line

34254

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Background of CT26-luc2-CEA6-CEA5-Cell-Line

CEACAM5 and CEACAM6, are also known as CD66e and CD66c. CEACAM5 is a cell surface glycoprotein that represents the founding member of the carcinoembryonic antigen (CEA) family of proteins. It takes part in cell adhesion and intracellular signaling as a cell surface glycoprotein, and also plays a role as an oncogene by promoting tumor progression, inducing resistance to anoikis of colorectal carcinoma cells. Furthermore, CEACAM5 mediates homophilic and heterophilic cell adhesion with other carcinoembryonic antigen-related cell adhesion molecules, such as CEACAM6. CEACAM6 is a cell surface glycoprotein that belongs to the CEA family whose members are glycosyl phosphatidyl inositol anchored cell surface glycoproteins. CEACAM6 plays a role in cell adhesion and is widely used as tumor markers in serum immunoassay determinations of carcinoma. CEACAM6 affects the sensitivity of tumor cells to adenovirus infection, and acts as a receptor for adherent-invasive E. coli adhesion to the surface of ileal epithelial cells in patients with Crohn's disease.

Specifications

Catalog Number	KC-3251
Cell Line Name	CT26-luc2-CEA6-CEA5-Cell-Line
Host Cell Line	CT26
Description	Stable CT26 cell line expressing exogenous luciferase under the control of CEACAM6-CEACAM5 signaling pathway.
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 + 600μg/mL Hygromycin B + 10μg/mL Puromycin
Selection Marker	Hygromycin B, Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1 : 4~1 : 8 every 3 days; seed out at about 1 × 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

CT26-Luc2-CEACAM6-CEACAM5 cell line was generated using a lentiviral vector expressing the luciferase, CEACAM5 and CEACAM6 sequence.

Characterization

Figure1: Characterization of CEACAM5 overexpression in CT26 stable clones using FACS.

Figure2: Characterization of CEACAM6 overexpression in CT26 stable clones using FACS.

Figure3: Characterization of CT26-Luc2-CEACAM6-CEACAM5 cell line stable clone using luciferase assay in the conditions of different cell numbers.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS, 600µg/mL Hygromycin B + 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 3 days; seed out at about 1 × 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

1. Entrez Gene: CEACAM6 carcinoembryonic antigen-related cell adhesion molecule 6 (non-specific cross reacting antigen).
2. Human PubMed Reference: National Center for Biotechnology Information, U.S. National Library of Medicine.
3. Benchimol S, Fuks A, Jothy S, Beauchemin N, Shirota K, Stanners CP. Carcinoembryonic antigen, a human tumor marker, functions as an intercellular adhesion molecule. Cell. 1989, 4, 57(2): 327–334.
4. Ilantzis C, DeMarte L, Screaton RA, Stanners CP. Deregulated expression of the human tumor marker CEA and CEA family member CEACAM6 disrupts tissue architecture and blocks colonocyte differentiation. Neoplasia. 4(2): 151–163.
5. Ordoñez C, Screaton RA, Ilantzis C, Stanners CP. Human carcinoembryonic antigen functions as a general inhibitor of anoikis. Cancer Research. 2000, July, 60 (13): 3419–3424.

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.