Esophageal Adenocarcinoma Methods and Protocols

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3 Methods

1. Before commencing cell culture, disinfect the hood cabinet
   with 70% ethanol. Disinfect gloves by spraying them with
   70% ethanol and allow to air dry before starting work (see
   Note 11).


2. Culture all esophageal adenocarcinoma cell lines (Table 1 )
   except Flo-1 in RPMI-1640 medium, whereas culture Flo-1 in
   DMEM medium supplemented with 10% fetal bovine serum,
   2 mM GlutaMax, 100 U/ml penicillin, and 100 mg/ml strep-
   tomycin at 37 °C in a humidified atmosphere of 95% air and
   5% CO 2 (see Note 3).


3. Expand cell number according to the need of your mice exper-
   iment using T175 flask. Harvest cells grown in monolayer cul-
   ture except ESO26 and ESO51 (which grow in suspension)
   during the exponential growth phase using trypsin-EDTA (see
   Note 19).


4. Before trypsinization, remove growth medium from cells and
   wash with 10 ml of DPBS. Aspirate DPBS, add 5 ml of
   trypsin- EDTA and incubate for 5 min, or until cells have
   detached, at 37 °C (see Note 20).

3.1 Preparation
of Cancer Cells

Table 1

Cell lines used to establish peritoneal dissemination xenograft mouse

models of esophageal adenocarcinoma

Cell lines name Cell lines origin

OE19 Adenocarcinoma of the esophagogastric junction

and gastric cardia (Caucasian)

OE33 Adenocarcinoma of the lower esophagus (with

Barrett metaplasia)

(Caucasian)

ESO26 Adenocarcinoma of the esophagogastric junction

and distal esophagus (Caucasian)

Flo-1 Adenocarcinoma of the distal esophagus

(Caucasian)

SK-GT-2 Adenocarcinoma of the gastric fundus (Hispanic

male), poorly differentiated

ESO51 Adenocarcinoma of the distal esophagus

(Caucasian) (Barrett metaplasia)

OACM5.1C Adenocarcinoma of the distal esophagus

(Caucasian)

Md Sazzad Hassan and Urs von Holzen