KC-5457

MKN45-CEACAM6-KO Cell Line

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Background of MKN45-CEACAM6-KO Cell Line

Carcinoembryonic antigen (CEA)-related cell adhesion molecule 6 (CEACAM6) is a cell adhesion protein of the CEA family of glycosyl phosphatidyl inositol anchored cell surface glycoproteins. It plays a significant role in cancer progression by inhibiting apoptosis, promoting drug resistance, and facilitating cancer cell invasion and metastasis. This protein is part of the carcinoembryonic antigen (CEA) family and plays a role in cell adhesion, proliferation, and tumor development. It is highly expressed in various cancers, including pancreatic, gastric, and colorectal cancers, making it a potential therapeutic target.

Specifications

Catalog Number	KC-5457
Cell Line Name	MKN45-CEACAM6-KO Cell Line
Clone Number	3A3
Host Cell Line	MKN45
Description	Stable MKN45 clone with human CEACAM6 gene knockout, No.3A3
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	RPMI1640+20% FBS+10% DMSO
Propagation Medium	RPMI1640+10% FBS
Selection Marker	NA
Morphology	Epithelial
Subculture	Split saturated culture 1:5-1:8 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 40 hours
Mycoplasma Status	Negative

Cell Line Generation

MKN45-CEACAM6-KO cell line was generated using the CRISPR method.

Characterization

Figure 1: Characterization of MKN45-CEACAM6-KO cell line stable clone using PCR sequencing.

Figure 2: Characterization of MKN45-CEACAM6-KO cell line stable clone using RT-PCR sequencing.

Figure 3: Characterization of MKN45-CEACAM6-KO cell line stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (RPMI1640 + 10% FBS) 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 1:5-1:8 every 2-3 days; seed out at about 1-3 ×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

Wu G, Wang D, Xiong F, Wang Q, Liu W, Chen J, Chen Y. The emerging roles of CEACAM6 in human cancer (Review). Int J Oncol. 2024 Mar;64(3):27. doi: 10.3892/ijo.2024.5615. Epub 2024 Jan 19. PMID: 38240103; PMCID: PMC10836497.
Zhao D, Cai F, Liu X, Li T, Zhao E, Wang X, Zheng Z. CEACAM6 expression and function in tumor biology: a comprehensive review. Discov Oncol. 2024 May 25;15(1):186. doi: 10.1007/s12672-024-01053-6. PMID: 38796667; PMCID: PMC11127906.