Pa tu 8902

The human pancreatic adenocarcinoma cell lines PANC-1, MiaPaCa-2, AsPC-1, BxPC-3, and PA-TU-8902 were purchased from the American Type Culture Collection. The pancreatic cancer cell line MDA Panc-28 was a gift from Dr. Paul J. Chiao (The University of Texas MD Anderson Cancer Center, Houston, TX). The human pancreatic adenocarcinoma cell line FG was obtained from Michael P. Vezeridis (The Warren Alpert Medical School of Brown University, Providence, RI) (16 (link)). The human metastatic pancreatic adenocarcinoma cell line COLO357 and its fast-growing liver-metastatic variant in nude mice, L3.7, were described previously (12 (link)). All of these cell lines were maintained in plastic flasks as adherent monolayers in Eagle's minimal essential medium supplemented with 10% fetal bovine serum (FBS), sodium pyruvate, nonessential amino acids, L-glutamine, and a vitamin solution (Flow Laboratories). PANC-1, MiaPaCa-2, AsPC-1, BxPC-3, and PA-TU-8902 were characterized or authenticated by the American Type Culture Collection using short tandem repeat profiling and passaged in our laboratory for fewer than 6 months after receipt. A competitive LDHA inhibitor, oxamate sodium, was purchased from Sigma-Aldrich (17 (link)).

All the human cell lines (Hs766T, BxPc3, Patu8988T and Patu8902) used in this study were purchased from ATCC, used below passage 25 and continuously cultured in 100 U/ml penicillin and 100 U/ml streptomycin. The cell lines were authenticated by short tandem repeat (STR) profiling at the Institute for Applied Cancer Sciences, MD Anderson Cancer Center. The Patu8988T, Hs766T and Patu8902 cell lines were routinely cultured in Dulbecco’s modified Eagle’s medium (DMEM) with 10% FBS (Invitrogen). BxPc3 cell lines were routinely cultured in Roswell Park Memorial Institute (RPMI) 1640 (Invitrogen) with 10% FBS. Primary mouse cell lines were established in the laboratory (AK-B6, AK192, HY6468, PJAK4217, PJAK4298) as described previously^{34 (link)} and were routinely cultured in Roswell Park Memorial Institute (RPMI) 1640 (Invitrogen) 10% FBS (Invitrogen). For inducible Kras derived cell lines, 1 ug/ml of doxycycline was directly added to the media. For metabolic and metabolomic assays, 10% dialysed FBS (Atlanta Biologicals Inc.) was used. The cell lines were mycoplasma free, based on tests done monthly in the laboratory using Lonza’s MycoAlert Mycoplasma Detection Kit assays with confirmatory tests by PCR-based assays.