KC-3231

PanO2-CDH17 Cell Line

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Background of PanO2-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-3231
Cell Line Name	PanO2-CDH17 Cell Line
Host Cell Line	PanO2
Description	Stable PanO2 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 + 2ug/ml Puromycin
Selection Marker	Puromycin
Morphology	Fibroblastoid cells growing as a monolayer
Subculture	Split saturated culture 1:4-1:5 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

PanO2 CDH17 cell line was generated using a lentiviral vector expressing the CDH17 sequence.

Characterization

Figure 1: Characterization of endogenous CDH17 expression in PanO2 cells using FACS.

Figure 2: Characterization of CDH17 overexpression in the PanO2 CDH17 stable clone using FACS.

Cell Resuscitation

1. Prewarm culture medium (RPMI1640 supplemented with 10% FBS and 2μg/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 1:4-1:5 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% RPMI1640 + 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.Yakirevich E, Magi-Galluzzi C, Grada Z, Lu S, Resnick MB, Mangray S. Cadherin 17 is a sensitive and specific marker for metanephric adenoma. Am J Surg Pathol. 2015 Apr;39(4):479-86. doi: 10.1097/PAS.0000000000000401. PMID: 25768256; PMCID: PMC4360917. 2. Bartolomé RA, Barderas R, Torres S, Fernandez-Aceñero MJ, Mendes M, García-Foncillas J, Lopez-Lucendo M, Casal JI. Cadherin-17 interacts with α2β1 integrin to regulate cell proliferation and adhesion in colorectal cancer cells causing liver metastasis. Oncogene. 2014 Mar 27;33(13):1658-69. doi: 10.1038/onc.2013.117. Epub 2013 Apr 22. PMID: 23604127.