KC-3118-DW

MC38-CDH17 Cell Line

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Home » 细胞系 » MC38-CDH17 Cell Line

Background of MC38-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 NumberKC-3118-DW
Cell Line NameMC38-CDH17 Cell Line
NCBI/UniProt Accession NumberNM_001144663.2
Clone NumberNA
Host Cell LineMC38
DescriptionStable MC38-CDH17 cell line expressing exogenous 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% DMEM+20% FBS+10% DMSO
Propagation MediumDMEM+10% FBS+5μg/mL Puromycin
Selection MarkerPuromycin
MorphologyEpithelial
SubcultureSplit saturated culture 1:4-1:10 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 20 hours
Mycoplasma StatusNegative

Cell Line Generation

MC38-CDH17 cell line was generated using lentivirus expressing CDH17 sequence.

Characterization

Figure 1. Characterization of CDH17 over-expression in the MC38-CDH17 stable clone using FACS.(Primary antibody: CDH17-hIgG1, Cat#KB-1062, Kyinno)

Figure 2:Validation of in vivo tumorigenicity of MC38 cells stably expressing CDH17 via subcutaneous implantation in C57BL/6J mice, with tumor growth monitored by measuring volume (V=0.5×length×width²) and body weight.

Cell Resuscitation

  1. Prewarm culture medium (DMEM+10% FBS+5μg/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 1:4-1:10 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. Entrez Gene: CDH17 cadherin 17, LI cadherin (liver-intestine).
  2. 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. doi:10.1006/geno.1998.5281. PMID 9615235.
  3. 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. doi:10.1006/geno.1998.5717. PMID 10191097.

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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