KC-6454

293T-cMet-KO-mouse-CDH17

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Background of 293T-cMet-KO-mouse-CDH17

CDH17 is a member of the cadherin superfamily, genes encoding calcium-dependent, membrane-associated glycoproteins. The encoded protein is cadherin-like, consisting of an extracellular region, containing 7 cadherin domains, and a transmembrane region but lacking the conserved cytoplasmic domain. The protein is a component of the gastrointestinal tract and pancreatic ducts, acting as an intestinal proton-dependent peptide transporter in the first step in oral absorption of many medically important peptide-based drugs. The protein may also play a role in the morphological organization of liver and intestine. Alternative splicing results in multiple transcript variants.

Specifications

Catalog Number	KC-6454
Cell Line Name	293T-cMet-KO-mouse-CDH17
NCBI/UniProt Accession Number	NM_019753.4
Clone Number	8#
Host Cell Line	293T
Description	Stable 293T-cMet-KO clone expressing exogenous mouse-CDH17 gene.
Quantity	One vial 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	DMEM+10%FBS+1μg/mL Puromycin
Selection Marker	Puromycin
Morphology	Human embryonic kidney cell line
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 24 hours
Mycoplasma Status	Negative

Cell Line Generation

293T-cMet-KO cell Line was generated using retrovirus vector expressing mouse-CDH17 sequence.

Characterization

Figure 1:Characterization of 293T-cMet-KO-mouse-CDH17 cell line stable clone using FACS.

Figure 2: Characterization of 293T-cMet-KO-mouse-CDH17 cell line stable clone using PCR sequencing.

Cell Resuscitation

1. Prewarm culture medium (DMEM supplemented with 10% FBS and 1μg/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 1:10 every 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

Liu X, Huang Y, Yuan H, Qi X, Manjunath Y, Avella D, Kaifi JT, Miao Y, Li M, Jiang K, Li G. Disruption of oncogenic liver-intestine cadherin (CDH17) drives apoptotic pancreatic cancer death. Cancer Lett. 2019 Jul 10;454:204-214. doi: 10.1016/j.canlet.2019.04.022. Epub 2019 Apr 17. PMID: 31004701.
Delaney S, Keinänen O, Lam D, Wolfe AL, Hamakubo T, Zeglis BM. Cadherin-17 as a target for the immunoPET of adenocarcinoma. Eur J Nucl Med Mol Imaging. 2024 Jul;51(9):2547-2557. doi: 10.1007/s00259-024-06709-7. Epub 2024 Apr 16. PMID: 38625402; PMCID: PMC11223962.

Use License Agreement

Research Use Only.

Not for use in diagnostic procedures or therapeutic applications.

Redistribution of the cell line or its derivatives is prohibited without prior written permission from Kyinno Biotechnology.