KC-3016-DW

CT26-CDH17 Cell Line

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Background of CT26-CDH17 Cell Line

CDH17 is a member of the cadherin superfamily, genes encoding calcium-dependent, membrane-associated glycoproteins. The protein is a component of the gastrointestinal tract and pancreatic ducts, is a highly specific marker for gastrointestinal adenocarcinoma and may be important in clinical diagnosis. The protein may also play a role in the morphological organization of liver and intestine.

Specifications

Catalog Number	KC-3016-DW
Cell Line Name	CT26-CDH17 Cell Line
Host Cell Line	CT26
Description	Stable CT26 clone expressing exogenous CDH17 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	Fibroblastoid
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

Cell Line Generation

CT26-CDH17 cell line was generated using a lentiviral vector expressing the CDH17 sequence.

Characterization

Figure 1: Characterization of CDH17 overexpression in the CT26-CDH17 stable clone using FACS.(Primary antibody: CDH17-hIgG1, Cat#KB-1062, Kyinno)

Figure 2: Validation of in vivo tumorigenicity of CT26 cells stably expressing CDH17 via subcutaneous implantation in BALB/c mice, with tumor growth monitored by measuring volume (V=0.5×length×width²) and body weight.

Cell Resuscitation

Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 10μg/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% RPMI1640 + 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

Altree-Tacha D, Tyrrell J, Haas T. CDH17 Is a More Sensitive Marker for Gastric Adenocarcinoma Than CK20 and CDX2. Arch Pathol Lab Med. 2017 Jan.
Yui A, Kuroda D, Maruno T, Nakakido M, Nagatoishi S, Uchiyama S, Tsumoto K. Molecular mechanism underlying the increased risk of colorectal cancer metastasis caused by single nucleotide polymorphisms in LI-cadherin gene. Sci Rep. 2023 Apr 20.