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
QuantityTwo vials 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.
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