Orthotopic Models of Esophageal Carcinoma and Their Use in Drug Discovery

Stephanie J. Gros1

1 Department of General, Visceral, and Thoracic Surgery, University Medical Center Hamburg‐Eppendorf, Hamburg, Germany
Publication Name:  Current Protocols in Pharmacology
Unit Number:  Unit 14.20
DOI:  10.1002/0471141755.ph1420s54
Online Posting Date:  September, 2011
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Abstract

The protocol detailed in this unit is for the establishment of an orthotopic model of human esophageal adenocarcinoma in NMRI/nu mice. The resultant tumor has high metastatic potential, spreading readily to liver, lungs, and lymph nodes. This model is useful for studying primary esophageal carcinoma, tumor biology, pathogenesis, tumor progression, metastatic homing, and the efficacy of therapeutic approaches for treating this condition. The practical use of this preclinical model for drug discovery is illustrated with data from a study on the chemotherapeutic effects of HER2‐targeted therapy. Curr. Protoc. Pharmacol. 54:14.20.1‐14.20.15. © 2011 by John Wiley & Sons, Inc.

Keywords: esophageal carcinoma; orthotopic implantation; metastatic potential; NMRI/nu mice; in vivo imaging

     
 
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Table of Contents

  • Commentary
  • Literature Cited
  • Figures
  • Tables
     
 
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Materials

Basic Protocol 1:

  Materials
  • Human cell line PT1590: isolated from a primary tumor of a patient with human esophageal adenocarcinoma at the University Medical Center Hamburg‐Eppendorf; it can be obtained upon request from the author ( )
  • Human cell line OE19: this cell line may be obtained from the European Collection of Cell Cultures, Health Protection Agency; Table 14.20.1 lists a sampling of esophageal cell lines and contacts that are available for research purposes
    Table 4.0.1   Materials   Esophageal Cell Lines a   Esophageal Cell Lines

    Cell line Cell type Source/literature reference
    OE19 AC Health Protection Culture Agency Collection, cat. no. 96071721
    OE33 AC Health Protection Culture Agency Collection, cat. no. 96070808
    OE21 SSC Health Protection Culture Agency Collection cat. no. 96062201
    KYSE‐150 SSC Health Protection Culture Agency Collection cat. no. 94072015
    KYSE‐270 SSC Health Protection Culture Agency Collection cat. no. 94072021
    KYSE‐30 SSC Health Protection Culture Agency Collection cat. no. 94072011
    KYSE‐410 SSC Health Protection Culture Agency Collection cat. no. 94072023
    KYSE‐70 SSC Health Protection Culture Agency Collection cat. no. 94072012
    JH‐EsoAd1 AC Alvarez et al. ( )
    OACP4.C AC de Both et al. ( )
    OACM4.1C AC de Both et al. ( )
    OACM5.1C AC de Both et al. ( )
    PT1590 AC Scheunemann et al. ( ), Gros et al. ( , b)
    SLMT‐1 SSC Tang et al. ( )
    HKESC‐1 SSC Hu et al. ( )
    ESC‐410 SSC Zhang et al. ( )

     aThis table gives a small sample of esophageal cell lines, but it is not a complete list. The author only has practical experience with cell lines OE19 and PT1590 regarding their use in the model.
  • RPMI 1690 medium (Linaris)
  • 10% fetal bovine serum (Linaris)
  • Penicillin/streptomycin (Biochrome KG)
  • Transferrin (Sigma‐Aldrich)
  • Insulin (Sigma‐Aldrich)
  • Basic fibroblast growth factor (Boehringer)
  • Epidermal growth factor (Boehringer)
  • 8‐ to 10‐week‐old (20 to 25 g) NMRI/nu (U.S. Naval Medical Research Institute; Charles River Deutschland) female mice
  • Tryspin
  • 1× PBS (Invitrogen)
  • Trypan blue
  • Ice
  • Surgical disinfectant or alcohol pads
  • Ketamine hydrochloride (Graeub)
  • Xylazine hydrochloride (Bayer)
  • Novamine sulfone (e.g., Ratiopharm)
  • Sterile gloves and surgical dress
  • Autoclave for sterilizing instruments
  • 5% CO 2 incubator
  • Operating microscope (Carl Zeiss or comparable product)
  • Cell counter (e.g., Neubauer‐counting chamber)
  • Centrifuge
  • 1‐ml syringe (BD Plastipak, Becton Dickinson)
  • 27‐G hypodermic needle (Sterican)
  • Animal scale
  • Calipers
  • Heating pad or heating lamp
  • Thermometer
  • Sterile surgical towels
  • Ear punch or tattooing device to identify individual mice (e.g., Fine Science Tools)
  • Petri dishes, 70‐mm or larger
  • Sterile surgical instruments (Fine Science Tools or comparable products) including:
    • Scalpel
    • Two pairs of scissors (tissue scissors and material scissor)
    • Two tissue forceps (Straight and bent)
    • Two needle holders (applicable for 8.0 prolene and 6.0 vicryl sutures)
    • Tissue spreader (e.g., Fine Science Tools)
  • 8.0 prolene suture (Ethicon or comparable product)
  • 6.0 vicryl sutures (Ethicon or comparable product)
  • Additional reagents and equipment for sacrificing the mice (Donovan and Brown, )
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Figures

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Literature Cited

Literature Cited
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