KC-3200

293T-CDH6 Cell Line

32141

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Background of 293T-CDH6 Cell Line

CDH6 (Cadherin-6), a type II calcium-dependent adhesion molecule, functions as an oncofetal antigen with dual roles in cancer. Normally restricted to embryonic kidney/thyroid development, it becomes aberrantly overexpressed (>60% cases) in clear cell renal cell carcinoma (ccRCC), ovarian cancer, and HCC through promoter demethylation. CDH6 promotes metastasis by activating Src/FAK signaling and disrupting E-cadherin-mediated adhesion, while simultaneously serving as a promising therapeutic target due to its exposed extracellular domains. Its clinical significance is highlighted as: (1) a diagnostic biomarker (urinary sCDH6 shows 82% sensitivity for early ccRCC), and (2) an emerging target for novel therapies – including CDH6-directed ADCs (e.g., HKT288 inducing complete responses in PDX models) and CAR-T cells showing efficacy in ovarian cancer preclinical studies. Notably, CDH6 exhibits tissue-context-dependent functions, demonstrating tumor-suppressive effects in papillary thyroid carcinoma, necessitating further investigation using engineered models to dissect its microenvironment-regulated signaling networks.

Specifications

Catalog Number	KC-3200
Cell Line Name	293T-CDH6 Cell Line
Host Cell Line	293T
Description	Stable 293T cell line expressing exogenous 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% 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	NA

Cell Line Generation

293T-CDH6 cell line was generated using lentivirus expressing CDH6 sequence.

Characterization

Figure 1. Characterization of CDH6 over-expression in the 293T-CDH6 stable clone using FACS.

Cell Resuscitation

Prewarm culture medium (DMEM supplemented with 10% FBS, 1μg/mL Puromycin)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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

Prepare the freezing medium (70% DMEM + 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

Shimazui T, Oosterwijk-Wakka J, Akaza H, Bringuier PP, Ruijter E, Debruyne FM, Schalken JA, Oosterwijk E. Alterations in expression of cadherin-6 and E-cadherin during kidney development and in renal cell carcinoma. Eur Urol. 2000 Sep;38(3):331-8. doi: 10.1159/000020302. PMID: 10940709.