R26 tdtomato

All animal studies and procedures were reviewed and approved by the Columbia University Medical Center Institutional Animal Care and Use Committee (IACUC), and all mice were bred under specific pathogen free (SPF) conditions. Cck2r-BAC-CreERT2-mice (Hayakawa et al., 2015b (link)) and Eef1a1-LSL-Notch1(IC) mice (Buonamici et al., 2009 (link)) have been previously reported. LSL-Trp53^R172H mice (Renz et al., 2018 (link)) were kindly provided by Dr. Kenneth Olive (Columbia University). Apc^flox/flox mice have been previously described (Westphalen et al., 2014 (link)) and were obtained from the National Cancer Institute (NCI). Lgr5-DTR-GFP mice, which have been previously reported (Asfaha et al., 2015 (link)), were provided by Genentech. Lgr5-GFP-IRES-CreERT2 mice (Lgr5-GFP mice) were provided by Dr. Hans Clevers. Gastrin-deficient (GAS-KO) mice (Koh et al., 1997 (link)) and Cck2r^−/− mice (Jin et al., 2009 (link)) have been reported in the past. R26-tdTomato and Cag-CreERT were purchased from the Jackson Laboratory. Cre recombinase was activated by oral administration of tamoxifen (2 mg/0.2 ml corn oil; Sigma-Aldrich).

To enable tracing of acinar lineage cells, an inducible acinar cell-specific reporter mouse was utilized. Briefly, the R26^tdTomato (Gt(ROSA)26Sortm9(CAG-tdTomato)Hze/J) reporter strain (Jackson Laboratory) was crossed to Mist1^CreERT2 on a C57/Bl6 background, as described previously (Aure, Konieczny, & Ovitt, 2015 (link)), to generate the Mist1^CreERT2;R26^TdTomato strain for acinar lineage tracing. Tamoxifen was dissolved in 10% ethanol and 90% corn oil at 60 mg/mL. Oral gavage with tamoxifen was performed for two consecutive days on 6–12 week old female mice, each at a dose of 0.25 mg/g, to induce Cre activation. SMG were harvested for culture one week following tamoxifen administration. Cell isolation and seeding were performed as previously described.

Mist1-CreERT2 mice (Shi et al., 2009 (link)), Cxcl12-dsRED mice (Ding and Morrison, 2013 (link)), Troy^−/− and Troy-BAC-CreERT2 mice (Fafilek et al., 2013 (link)), H/K-ATPase-Cxcl12 mice (Shibata et al., 2013 (link)), Eef1a1-LSL-Notch1(IC) mice (Buonamici et al., 2009 (link)), Wnt5a^flox mice (Miyoshi et al., 2012 (link)) were described previously. Cxcr4-EGFP mice were kindly provided by Richard J. Miller (Northwestern University Medical School, USA). LSL-Kras^G12D and LSL-Trp53^R172H mice were provided by Dr. Kenneth Olive (Columbia University, USA). Apc^flox mice were obtained from the National Cancer Institute (NCI). Lgr5-DTR-GFP mice were provided by Genentech. Cdh1^flox, R26-mTmG, R26-LacZ, R26-TdTomato, R26-Confetti, R26-EYFP, Cxcl12^flox, Tie2-Cre, Id2-GFP, and Cag-CreERT2 mice were purchased from the Jackson Laboratory. Cre recombinase was activated by oral administration of TAM (1–5mg/0.2mL corn oil, as indicated). All animal studies and procedures were approved by the ethics committees at Columbia University and the Academy of Sciences of the Czech Republic. Human stomach tissue sections were obtained from DGC patients who underwent surgical resection or endoscopic submucosal dissection from 2001 to 2012 at Gifu University Hospital, Gifu, Japan. All study protocols were approved by the ethics committees, and written informed consent was obtained from all patients.