KC-3429

CHOK1-cyno-CDH17-Middle Cell Line

34382

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Background of CHOK1-cyno-CDH17-Middle Cell Line

CDH17,also known as HPT1, this gene 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-3429
Cell Line Name	CHOK1-cyno-CDH17-Middle Cell Line
NCBI/UniProt Accession Number	XM_045398503.1
Clone Number	NA
Host Cell Line	CHOK1
Description	Stable CHOK1 cell line expressing exogenous cyno-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	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
In Vivo Validation	NA

Cell Line Generation

CHOK1-cyno-CDH17-Middle cell line was generated using lentivirus expressing cyno-CDH17 sequence.

Characterization

Figure 1. Characterization of cyno-CDH17 over-expression in the CHOK1-cyno-CDH17-Middle stable clone using FACS.(Primary antibody: CDH17-hIgG1, Cat#KB-1062, Kyinno)

Cell Resuscitation

Prewarm culture medium (RPMI1640+10% FBS +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

1.Chang YY, Yu LC, Yu IS, Jhuang YL, Huang WJ, Yang CY, Jeng YM. Deletion of cadherin-17 enhances intestinal permeability and susceptibility to intestinal tumour formation. J Pathol. 2018 Nov;246(3):289-299.
Okada T, Kurabayashi A, Akimitsu N, Furihata M. Expression of Cadherin-17 Promotes Metastasis in a Highly Bone Marrow Metastatic Murine Breast Cancer Model. Biomed Res Int. 2017;2017:8494286.
Funakoshi S, Shimizu T, Numata O, Ato M, Melchers F, Ohnishi K. BILL-cadherin/cadherin-17 contributes to the survival of memory B cells. PLoS One. 2015 Jan 22;10(1):e0117566.