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KC-5456

MKN45-CEACAM6-KO Cell Line

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Datasheet
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Home » MKN45-CEACAM6-KO Cell Line

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 NumberKC-5456
Cell Line NameMKN45-CEACAM6-KO Cell Line
Clone Number1C2
Host Cell LineMKN45
DescriptionStable MKN45 clone with human CEACAM6 gene knockout, No.1C2
QuantityOne vial of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing MediumRPMI1640+20% FBS+10% DMSO
Propagation MediumRPMI1640+10% FBS
Selection MarkerNA
MorphologyEpithelial
SubcultureSplit saturated culture 1:5-1:8 every 2-3 days; seed out at about 1-3 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 40 hours
Mycoplasma StatusNegative

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

  1. Prewarm culture medium (RPMI1640 + 10% FBS) 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 1:5-1:8 every 2-3 days; seed out at about 1-3 ×105 cells/mL.

Cell Freezing

  1. Prepare the freezing medium (70% RPMI1640 + 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. 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.
  2. 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.

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.

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