KC-3176

293T-cyno-CDH6-Cell-Line

Stable HEK293T cell line expressing exogenous cyno CDH6 gene

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Background of 293T-cyno-CDH6-Cell-Line

CDH6, also known as CAD6, is a type II classical cadherin that belongs to the cadherin superfamily. CDH6 has high expressions in kidney, brain, and cerebellum, and low expressions in lung, pancreas, gastric mucosa, and cytotrophoblasts. CDH6 is also found in renal, lung, and ovarian carcinoma. As a classic cadherin, CDH6 will form homodimers and promote intercellular adhesion with itself and possibly, cadherin-9 and -14. Decreased expression of CDH6 may be associated with tumor growth and metastasis.

Specifications

Catalog Number	KC-3176
Cell Line Name	293T-cyno-CDH6-Cell-Line
Description	Stable HEK293T cell line expressing exogenous cyno 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 1:3-1:6 every 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 cyno CDH6 Cell Line was generated using a lentiviral vector expressing the cyno CDH6 sequence.

Characterization

Cell Resuscitation

Prewarm culture medium (DMEM + 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 1:3-1:6 every 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

Shimoyama, Y. et al. (2000) Biochem. J. 349:159.
Shimoyama, Y. et al. (1999) J. Biol. Chem. 274:11987.
Shimoyama Y, Gotoh M, Terasaki T, Kitajima M, Hirohashi S (June 1995). "Isolation and sequence analysis of human cadherin-6 complementary DNA for the full coding sequence and its expression in human carcinoma cells". Cancer Res. 55 (10): 2206ÿ11. PMID 7743525.
Chalmers IJ, Hofler H, Atkinson MJ (June 1999). "Mapping of a cadherin gene cluster to a region of chromosome 5 subject to frequent allelic loss in carcinoma". Genomics. 57 (1): 160ÿ3. doi:10.1006/geno.1998.5717. PMID 10191097.
Entrez Gene: CDH6 cadherin 6, type 2, K-cadherin (fetal kidney).