KC-2123

MC38-CD66e-Cell-Line

23436

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Background of MC38-CD66e-Cell-Line

CD66e, also named CEA, is a glycoprotein that belongs to the large family of CEACAM and pregnancy specific glycoproteins, which is plays a role in cell adhesion and in intracellular signaling. Multiple cellular activities have been attributed to the encoded protein, including roles in the differentiation and arrangement of tissue threedimensional structure, angiogenesis, apoptosis, tumor suppression, metastasis, and the modulation of innate and adaptive immune responses. CD66e is one of the most widely used tumor markers in serum immunoassay determinations of carcinoma.

Specifications

Catalog Number	KC-2123
Cell Line Name	MC38-CD66e-Cell-Line
Host Cell Line	Mouse MC38 cell line
Description	Stable MC38 clone expressing exogenous human CD66e 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+5µg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:2-1:5 every 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 48 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

MC38 human CD66e Cell Line was generated using a retroviral vector expressing human CD66e sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS + 5µg/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 1:2-1:5 every 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

Beauchemin N, Arabzadeh A (December 2013). "Carcinoembryonic antigen-related cell adhesion molecules (CEACAMs) in cancer progression and metastasis". Cancer and Metastasis Reviews. 32 (3ÿ4): 643ÿ671.
Benchimol, Sarita; Fuks, Abraham; Jothy, Serge; Beauchemin, Nicole; Shirota, Kinji; Stanners, Clifford P. (April 1989). "Carcinoembryonic antigen, a human tumor marker, functions as an intercellular adhesion molecule". Cell. 57 (2): 327ÿ334. doi:10.1016/0092-8674(89)90970-7.
Ilantzis C, DeMarte L, Screaton RA, Stanners CP (2002). "Deregulated expression of the human tumor marker CEA and CEA family member CEACAM6 disrupts tissue architecture and blocks colonocyte differentiation". Neoplasia.
4 (2): 151ÿ163. doi:10.1038/sj.neo.7900201. PMC 1550325. PMID 11896570. 4. Ordo?ez C, Screaton RA, Ilantzis C, Stanners CP (July 2000). "Human carcinoembryonic antigen functions as a general inhibitor of anoikis". Cancer Research. 60 (13): 3419ÿ3424. PMID 10910050