KC-5451

CHOK1-cyno-CDH1-Cell-Line

63999

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Background of CHOK1-cyno-CDH1-Cell-Line

CDH1 (Cadherin 1) is a Protein Coding gene, it is encodes a classical cadherin of the cadherin superfamily. E-Cadherin protein is necessary for physiological signaling pathways, such as cell proliferation, maintenance of cell adhesion, cell polarity and epithelial-mesenchymal transition. it dysregulation leads to tumor proliferation, invasion, migration and metastases. Germline mutation in CDH1 (E-cadherin) tumor suppressor gene is associated with hereditary diffuse gastric cancer (HDGC) and lobular breast cancers (LBC). Diseases associated with CDH1 include Diffuse Gastric And Lobular Breast Cancer Syndrome and Blepharocheilodontic Syndrome 1.

Specifications

Catalog Number	KC-5451
Cell Line Name	CHOK1-cyno-CDH1-Cell-Line
NCBI/UniProt Accession Number	XM_005592359.5
Clone Number	3#
Host Cell Line	CHOK1
Description	Stable CHOK1 clone expressing exogenous cyno-CDH1 gene.
Quantity	One vial 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% RPMI 1640 + 20% FBS + 10% DMSO
Propagation Medium	RPMI 1640 + 10% FBS + 10μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Epithelial
Subculture	Split saturated culture 1:6-1:8 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 24 hours
Mycoplasma Status	Negative

Cell Line Generation

CHOK1-cyno-CDH1-cell-line was generated using a lentiviral vector expressing the cyno-CDH1 sequence.

Characterization

Figure 1: Characterization of cyno-CDH1 overexpression in the CHOK1-cyno-CDH1 stable clone using FACS.

Figure 2: Characterization of cyno-CDH1 in the CHOK1-cyno-CDH1 stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (RPMI 1640 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 1:6-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI 1640 + 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.Wang, J., Xiu, J., Battaglin, F. et al. Large-scale analysis of CDH1 mutations defines a distinctive molecular subset with treatment implications in gastric cancer. npj Precis. Onc. 8, 214 (2024).
2.Shenoy S. CDH1 (E-Cadherin) Mutation and Gastric Cancer: Genetics, Molecular Mechanisms and Guidelines for Management. Cancer Manag Res. 2019 Dec 13;11:10477-10486. doi: 10.2147/CMAR.S208818. PMID: 31853199; PMCID: PMC6916690.
3.Adib, E., El Zarif, T., Nassar, A.H. et al. CDH1 germline variants are enriched in patients with colorectal cancer, gastric cancer, and breast cancer. Br J Cancer 126, 797–803 (2022).

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.