KC-3716

CHOK1-rat-CDH6

34652

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Background of CHOK1-rat-CDH6

CDH6 (cadherin-6 or K-cadherin) is a single-transmembrane protein belonging to the Type II classical cadherin family, consisting of 790 amino acids, including extracellular domains, transmembrane regions, and intracellular tails. Its genes are located on human chromosome 5 and play an important role mainly in the basal outer membrane of epithelial cells, responsible for mediating calcium-dependent intercellular adhesion, maintaining tissue integrity and normal cell arrangement.

Specifications

Catalog Number	KC-3716
Cell Line Name	CHOK1-rat-CDH6
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous rat-CDH6 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 + 10μg/ml Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:4-1:8 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 30 hours
Mycoplasma Status	Negative
In Vivo Validation	NA

Cell Line Generation

CHOK1-rat-CDH6 cell line was generated using a lentiviral vector expressing the rat-CDH6 sequence.

Characterization

Figure 1: Characterization of rat CDH6 overexpression in the CHOK1 rat CDH6 stable clone using FACS.

Figure 2: Characterization of rat CDH6 overexpression in the CHOK1 rat CDH6 stable clone using PCR sequence.

Cell Resuscitation

1. Prewarm culture medium (70% RPMI1640 supplemented with 10% FBS and 10μg/mL puromycin)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 1:4-1:8 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. Suzuki H, Nagase S, Saito C, Takatsuka A, Nagata M, Honda K, Kaneda Y, Nishiya Y, Honda T, Ishizaka T, Nakamura K, Nakada T, Abe Y, Agatsuma T. Raludotatug Deruxtecan, a CDH6-Targeting Antibody-Drug Conjugate with a DNA Topoisomerase I Inhibitor DXd, Is Efficacious in Human Ovarian and Kidney Cancer Models. Mol Cancer Ther. 2024 Mar 4;23(3):257-271. doi: 10.1158/1535-7163.MCT-23-0287. PMID: 38205802; PMCID: PMC10911705. 2. Meng M, Zhou H, He Y, Chen L, Wang W, Yang L, Wang Z, Zhang L, Wang S. CDH6 as a prognostic indicator and marker for chemotherapy in gliomas. Front Genet. 2022 Jul 22;13:949552. doi: 10.3389/fgene.2022.949552. PMID: 35938030; PMCID: PMC9355303. 3. Ahmad S, Milan MDC, Hansson O, Demirkan A, Agustin R, Sáez ME, Giagtzoglou N, Cabrera-Socorro A, Bakker MHM, Ramirez A, Hankemeier T, Stomrud E, Mattsson- Carlgren N, Scheltens P, van der Flier WM, Ikram MA, Malarstig A, Teunissen CE, Amin N, van Duijn CM. CDH6 and HAGH protein levels in plasma associate with Alzheimer's disease in APOE ε4 carriers. Sci Rep. 2020 May 19;10(1):8233. doi: 10.1038/s41598-020-65038-5. PMID: 32427856; PMCID: PMC7237496.