KC-3390

CHOK1-mouse-CDH17 Cell Line

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Home » 细胞系 » CHOK1-mouse-CDH17 Cell Line

Background of CHOK1-mouse-CDH17 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 NumberKC-3390
Cell Line NameCHOK1-mouse-CDH17 Cell Line
NCBI/UniProt Accession NumberNM_019753.4
Clone Number3#
Host Cell LineCHOK1
DescriptionStable CHOK1 cell line expressing exogenous mouse-CDH17 gene
QuantityOne vial of frozen cells (≥2-106/vial)
StabilityStable in culture over a minimum of 10 passages
ApplicationDrug screening and biological assays
Freezing Medium70% RPMI1640+20% FBS+10% DMSO
Propagation MediumRPMI1640+10% FBS +10μg/mL Puromycin
Selection MarkerPuromycin
MorphologyEpithelial
SubcultureSplit saturated culture 1:4-1:8 every 2-3 days; seed out at about 1-3 × 105 cells/mL
Incubation37 °C with 5% CO2
StorageLiquid nitrogen immediately upon receiving
Doubling TimeApproximately 30 hours
Mycoplasma StatusNegative
In Vivo ValidationNA

Cell Line Generation

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

Characterization

Figure 1. Characterization of mouse-CDH17 over-expression in the CHOK1-mouse-CDH17 stable clone using qPCR

Figure 2. Characterization of mouse-CDH17 over-expression in the CHOK1-mouse-CDH17 stable clone using PCR

Cell Resuscitation

  1. Prewarm culture medium (RPMI1640+10% FBS +10μg/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% 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. 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.
  2. 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.
  3. 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.

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.
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