KC-3440

293T-Rat-CDH17 Cell Line

Stable HEK293T cell line expressing exogenous rat CDH17 gene

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34390
Home » 细胞系 » 293T-Rat-CDH17 Cell Line

Background of 293T-Rat-CDH17 Cell Line

CDH17 is a member of the cadherin superfamily, genes encoding calcium-dependent, membrane-associated glycoproteins. The protein is a component of the gastrointestinal tract and pancreatic ducts, is a highly specific marker for gastrointestinal adenocarcinoma and may be important in clinical diagnosis. The protein may also play a role in the morphological organization of liver and intestine.

Specifications

Catalog NumberKC-3440
Cell Line Name293T-Rat-CDH17 Cell Line
Clone Number5#
Host Cell Line293T
DescriptionStable 293T cell line expressing exogenous rat 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 + 1μg/mL Puromycin
Selection MarkerPuromycin
MorphologyEpithelial
SubcultureSplit saturated culture 1:4-1:5 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

293T-Rat-CDH17 cell line was generated using a lentiviral vector expressing the rat CDH17 sequence.

Characterization

Figure 1: Characterization of rat CDH17 overexpression in the 293T Rat CDH17 stable clone using QPCR.

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 1:4-1:5 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. Altree-Tacha D, Tyrrell J, Haas T. CDH17 Is a More Sensitive Marker for Gastric Adenocarcinoma Than CK20 and CDX2. Arch Pathol Lab Med. 2017 Jan.
  2. Sommerfeld, Mark R, Günter Müller, Georg Tschank, Gerhard Seipke, Paul Habermann, Roland Kurrle, and Norbert Tennagels. 2010. “In Vitro Metabolic and Mitogenic Signaling of Insulin Glargine and Its Metabolites.” Edited by Kathrin Maedler. PLoS ONE 5 (3): e9540–49.
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