KC-3748

293T-CDH3 cell line

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Background of 293T-CDH3 cell line

CDH1 encodes the classical cadherin of the cadherin superfamily. Alternative splicing leads to multiple transcript variants, at least one of which encodes a proprotein that is proteolytically processed to produce the mature glycoprotein. This calcium-dependent cell adhesion protein consists of five extracellular cadherin repeats, a transmembrane region, and a highly conserved cytoplasmic tail. This gene is located in a cluster of genes on the long arm of chromosome 16, a region associated with loss of heterozygosity events in breast and prostate cancers. Additionally, abnormal expression of this protein has been observed in cervical adenocarcinoma.

Specifications

Catalog Number	KC-3748
Cell Line Name	293T-CDH3 cell line
Host Cell Line	293T
Description	Stable HEK293T clone expressing exogenous CDH3 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% DMEM + 20% FBS + 10% DMSO
Propagation Medium	DMEM + 10% FBS + 1μg/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	Yes

Cell Line Generation

293T CDH3 cell line was generated using lentiviral vector expressing CDH3 sequence

Characterization

Figure 1: Characterization of CDH3 overexpression in the 293T CDH3 stable clone using FACS.

Figure 2: Characterization of human CDH3 overexpression in the 293T human CDH3 stable clone using PCR sequence

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1ug/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 x 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 x 10⁵ cells/ml.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 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 x g for 5 minutes at room temperature and carefully aspirate off the medium.
5. Resuspend the cells at a density of at least 3 x10⁶cells/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. Zhou Y, Chi Y, Bhandari A, Xia E, Thakur PC, Qu J, Wang O, Zhang X.Downregulated CDH3 decreases proliferation, migration, and invasion in thyroid cancer. Am J Transl Res. 2020 Jun 15;12(6):3057-3067. PMID: 32655830; PMCID:PMC7344063.
2. Wu T, Xiao Z, Li Y, Jiao Z, Liang X, Zhang Y, Liu H, Yang A. CDH3 is associated with poor prognosis by promoting the malignancy and chemoresistance in oral squamous cell carcinoma. Asian J Surg. 2022 Dec;45(12):2651-2658. doi: 10.1016/j.asjsur.2022.01.075. Epub 2022 Mar 17. PMID: 35305877.
3.Satavisa S, Padhy SK. CDH3 Mutation-Associated Juvenile-Onset Macular Dystrophy. Ophthalmol Retina. 2025 Feb;9(2):e12. doi:10.1016/j.oret.2024.06.016. Epub 2024 Jul 23. PMID: 39046397.