KC-6567

293T-cMet-KO-cyno-CDH17-Cell-Line

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Background of 293T-cMet-KO-cyno-CDH17-Cell-Line

CDH17, also known as HPT1, is a protein that belongs to the cadherin family of calcium-dependent cell adhesion molecules. CDH17 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. CDH17 may also play a role in the morphological organization of liver and intestine.

Specifications

Catalog Number	KC-6567
Cell Line Name	293T-cMet-KO-cyno-CDH17-Cell-Line
NCBI/UniProt Accession Number	XM_045398503.1
Clone Number	14#
Host Cell Line	293T-cMet-KO
Description	Stable 293T-cMet-KO clone expressing exogenous cyno-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% 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

Cell Line Generation

293T-cMet-KO-cyno-CDH17-cell-line was generated using a lentiviral vector expressing the cyno-CDH17 sequence.

Characterization

Figure 1: Characterization of cyno-CDH17 overexpression in the 293T-cMet-KO-cyno-CDH17 stable clone using FACS.

Figure 2: Characterization of cyno-CDH17 in the 293T-cMet-KO-cyno-CDH17 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 1:4-1:8 every 2-3 days; seed out at about 1-3 × 10⁵ cells/mL.

Cell Freezing

1. Prepare the freezing medium (70% DMEM + 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. Kremmidiotis G, Baker E, Crawford J, Eyre HJ, Nahmias J, Callen DF (Aug 1998). "Localization of human cadherin genes to chromosome regions exhibiting cancer-related loss of heterozygosity". Genomics. 49 (3): 467–71.
2.Chalmers IJ, Hofler H, Atkinson MJ (Jun 1999). "Mapping of a cadherin gene cluster to a region of chromosome 5 subject to frequent allelic loss in carcinoma". Genomics. 57 (1): 160–3.

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.