KC-3662

293T-CEACAM6 Cell Line

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Background of 293T-CEACAM6 Cell Line

CEACAM6 encodes CEA cell adhesion molecule 6 and is also known as NCA. Members of this family play a role in cell adhesion and are widely used as tumor markers in serum immunoassay determinations of carcinoma. This gene affects the sensitivity of tumor cells to adenovirus infection. The protein encoded by this gene acts as a receptor for adherent-invasive E. coli adhesion to the surface of ileal epithelial cells in patients with Crohn's disease. This gene is clustered with genes and pseudogenes of the cell adhesion molecules subgroup of the CEA family on chromosome 19.

Specifications

Catalog NumberKC-3662
Cell Line Name293T-CEACAM6 Cell Line
Host Cell Line293T
Description293T-CEACAM6 is a stable engineered cell line expressing human CEACAM6
QuantityOne vial of frozen cells (≥2×10⁶ cells/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% DMEM+20% FBS+10% DMSO
Propagation MediumDMEM+10%FBS+1ug/mL Puromycin
Selection MarkerPuromycin
MorphologyEpithelial
SubcultureSplit at a ratio of 1:4–1:6 every 2–3 days
Incubation37 °C with 5% CO₂
StorageLiquid nitrogen immediately upon receiving
Doubling TimeEstimated 19–36 hours
Mycoplasma StatusNegative
In Vivo ValidationNA

Cell Line Generation

293T-CEACAM6 cell line was generated using a lentiviral vector expressing the human CEACAM6 sequence.

Characterization

Figure 1: Characterization of CEACAM6 overexpression in the 293T CEACAM6 stable clone using FACS.

Figure 2: Characterization of CEACAM6 overexpression in 293T cell using FACS.

Cell Resuscitation

  1. Pre-warm complete culture medium (basal medium and 10% FBS) in a 37°C water bath.
  2. Rapidly thaw the cryovial in a 37°C water bath for 1-2 minutes with gentle agitation.
  3. Transfer the vial to a biosafety cabinet, and disinfect the exterior with 70% ethanol.
  4. Aseptically transfer the cell suspension dropwise into a sterile centrifuge tube containing 9.0 mL of pre-warmed complete medium.
  5. Centrifuge at approximately 125 × g for 5–7 minutes at room temperature, carefully aspirate the supernatant without disturbing the cell pellet.
  6. Gently resuspend the pellet in an appropriate volume of complete medium and transfer the suspension into a T25 flask.
  7. Incubate the flask in a 37°C in a humidified 5% CO₂ incubator.
  8. Assess cell viability and morphology after 24 hours. If cells appear healthy, replace the medium with fresh medium supplemented with the appropriate selective antibiotic.
  9. Subculture the cells at a ratio of 1:4–1:6 every 2–3 days upon reaching 80%–90% confluency.

Cell Freezing

  1. Prepare the freezing medium (70% basal medium, 20% FBS and 10% DMSO) freshly before use.
  2. Pre-chill the freezing medium on ice and label the cryovials accordingly.
  3. Transfer the cell suspension to a sterile conical tube and perform a cell count to determine total viability and density.
  4. Centrifuge the cells at 250×g for 5 minutes at room temperature; carefully aspirate the supernatant.
  5. Gently resuspend the pellet in chilled freezing medium, ensuring a minimum cell density of 3×10⁶ cells/mL.
  6. Aliquot 1 mL of the cell suspension into each pre-labeled cryovial.
  7. Place the cryovials into a CoolCell® container and store at -80°C overnight for controlled-rate cooling.
  8. Transfer the cryovials to the liquid nitrogen for long-term storage the following day.

References

  1. Piejko M, Bak K, Wierciak J et al. Targeting Circulating Tumor Cells in Pancreatic Ductal Adenocarcinoma: Rationale, Current Evidence, and a CEACAM6 CAR-T Strategy. Cancers. 2026 Jun 5;18(11):. PMID: 42279434. doi: 10.3390/cancers18111852.
  2. Al-Khinji A Exploring the interactions of integrins and CEACAM6 (Review). Experimental and therapeutic medicine. 2025 Dec;30(6):237. PMID: 41089193. doi: 10.3892/etm.2025.12986.
  3. Zhao D, Cai F, Liu X et al. CEACAM6 expression and function in tumor biology: a comprehensive review. Discover oncology. 2024 May 25;15(1):186. PMID: 38796667. doi: 10.1007/s12672-024-01053-6.

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.