Esophageal Adenocarcinoma Methods and Protocols

153

of the major obstacles in developing novel therapies for patients

with esophageal adenocarcinoma has been the lack of an animal

survival model for testing these anticancer pharmacotherapeutics.

Thus, improvement of outcomes of patients with esophageal ade-

nocarcinoma requires well-characterized animal survival models in

which to evaluate novel therapeutics.

Here we describe a subcutaneous mouse xenograft model as

well as a peritoneal dissemination mouse xenograft model for sur-

vival outcome analysis with intraperitoneal injection of human

esophageal adenocarcinoma cell lines. Mice survival studies were

then performed with clinically proven anticancer drugs paclitaxel

and carboplatin treatments.

2 Materials

Non-obese-diabetic/severe combined immune deficient mice

(NOD/SCID) mice, female, 4–6 weeks old (see Note 1).

Human esophageal adenocarcinoma cell lines (see Note 2):

OE19; OE33; ESO26; Flo-1; SK-GT-2; ESO51; OACM5.1C.

1. 
   

   RPMI-1640 medium (see Note 3).

   
   


2. 
   

   DMEM medium (see Note 3).

   
   


3. 
   

   Fetal bovine serum (see Note 4).

   
   


4. 
   

   GlutaMax 100× (see Note 5).

   
   


5. 
   

   Penicillin/Streptomycin solution 100×, 5000 U/ml (see
   Note 6 ).

   
   


6. 
   

   Dulbecco’s phosphate buffered saline (DPBS) (see Note 7).

   
   


7. 
   

   Trypsin-EDTA (0.25%), phenol red (see Note 8).

   
   


8. 
   

   Trypan blue (see Note 9).

   
   


9. 
   

   Paclitaxel (see Note 10).

   
   


10. 
    

    Carboplatin (see Note 10).

    
    


11. 
    

    Biological safety cabinet (see Note 11).

    
    


12. 
    

    1 ml Tuberculin syringe (see Note 12).

    
    


13. 
    

    Needle 25 G × (^1) 1/2 (0.5 mm × 40 mm) (see Note 13).

    
    


14. 
    

    Needle 25 G × 5/8 (0.5 mm × 16 mm) (see Note 14).

    
    


15. 
    

    Needle 27 G × 1/2 (0.4 mm × 13 mm) (see Note 15).

    
    


16. 
    

    Needle 30 G × 1/2 (0.3 mm × 13 mm) (see Note 16).

    
    


17. 
    

    Centrifuge (see Note 17).

    
    


18. 
    

    Hemocytometer (see Note 18).

    
    


19. 
    

    Microscope (see Note 18).

    
    

2.1 Mice

2.2 Cancer Cell Lines
(See Table 1 ) [ 12 ]

2.3 Media, Solutions,
and Reagents

2.4 Equipment

Xenograft Models of Esophageal Adenocarcinoma