Esophageal Adenocarcinoma Methods and Protocols

158

0

0

50

Cumulative survival

100

20

(^4653)
57
ControlCarboplatin∗
Median survival (Days)
Paclitaxel∗
40 60
Time (Days)
Fig. 3 Mice survival in the OE19 peritoneal dissemination model treated with
carboplatin and paclitaxel. 10 × 106 OE19 cells were injected intraperitoneally in
SCID mice and treatment started after 2 weeks and continued for another
2 weeks. The curve represents the animal survival time from the day of implan-
tation. Asterisk represents significant differences compared with control (vehicle)
at p = 0.0034 (published in PLoS One. 2017 Jun 19;12(6):e0180146 and permis-
sion obtained from publisher to reuse)

6. After euthanasia, examine the mouse for the presence and
   extent of intra-abdominal tumors. Harvest the peritoneal
   tumors and hepatic implants (see Note 34) and immerse them
   in 4% formaldehyde for fixation followed by paraffin embed-
   ding for histology (Fig. 4 ) [ 12 ].

4 Notes

1. Choosing SCID mice over athymic nude mice for the meta-
   static survival effect is beneficial because of more reproducibil-
   ity of the effect observed in SCID mice over nude mice [ 12 ].
   We chose female mice because they are easier to handle. The
   ideal age of mice should be between 4 and 6 weeks so that we
   have some time window for therapeutic interventions as well as
   to see survival benefit.


2. Choosing of appropriate esophageal adenocarcinoma cell lines
   should be done carefully as mistaken identity has been observed
   for widely used esophageal adenocarcinoma cell lines [ 30 ].


3. Use of appropriate media and cell culture conditions should be
   followed according to the American Type Culture Collection
   (ATCC) cell culture guidelines for specific cell types.


4. Use heat inactivation of fetal bovine serum (FBS) by heating it
   for 30 min at 56 °C with mixing to activate complement.

Md Sazzad Hassan and Urs von Holzen