Mycaway

The primary human PDAC cell lines PANC-1 and MIA PaCa₂ were cultured in high glucose DMEM (HyClone, Logan, UT, United States), while the metastatic SW1990 and ASPC-1 lines (all from American Type Culture Collection, ATCC, Manassas, VA, United States) were cultured in RPMI1640 medium (HyClone, Logan, UT, United States). Both media were supplemented with 10% heat-inactivated FBS (HyClone, Logan, UT, United States), penicillin-streptomycin solution (Gibco, CA, United States), and 2.5% horse serum (Gibco, Carlsbad, CA, United States) was included for MIA PaCa₂. All cell lines were cultured at 37 °C under 5% CO₂. The cells were routinely tested using MycAway (Yeasen, Shanghai, China) to eliminate mycoplasma contamination. Paired tumor and adjacent normal tissue samples were collected from the Shanghai General Hospital, and informed consent was obtained from the patients. The study was approved by the Ethics Committee of Shanghai General Hospital of Shanghai Jiao Tong University.

A human normal colorectal mucosal cell line (FHC) and CRC cell lines (HT29, HCT116, LOVO, SW480 and SW620) were bought from the American Type Culture Collection (ATCC, Manassas, VA, United States). All cells were cultured in Roswell Park Memorial Institute (RPMI) 1640 medium (Invitrogen, Carlsbad, CA, United States) containing 10% fetal bovine serum (FBS) (Gibco, Gaithersburg, MD, United States) and 1% penicillin/streptomycin (Gibco), which were incubated at 37 °C under 5% CO₂. To eliminate mycoplasma contamination, all cells were routinely examined using MycAway (Yeasen, Shanghai, China).