Pancreatic Cell Lines and Islet Isolation

Human pancreatic cell lines PANC-1 and pancreatic β-cell lines INS-1 were purchased from The Cell Bank of Type Culture Collection of Chinese Academy of Sciences (Shanghai, China). Islets were isolated from healthy rats and model rats and dispersed into single cells according to an earlier report [38 (link)]. PANC-1 cells were cultured in DMEM, and INS-1 and isolated islet cells were cultured in RPMI 1640 with 10% (v/v) FCS, 10 mM HEPES and 2 mM glutamine, 100 μg/mL penicillin, and 100 μg/mL streptomycin at 37°C and 5% CO₂. Pancreatic cancer cell line PANC-1 was transfected with the plasmids ptfLC3 (an indicator of autophagic activity, Addgene; ID 21074, Cambridge, MA, USA), pcDNA3.1-NF-κB, and pTZU6+1-shRNA-NF-κB. Transfection was performed via Lipofectamine 2000TM (Invitrogen, Carlsbad, CA, USA). After a two-day transfection, the antibiotic-resistant cells were collected by using 200 μg/mL G-418. The strain was cultured at the same condition for three days. 25 mg/L picroside II was added to PANC-1, INS-1, and isolated islet cells and cultured in DMEM at 37°C and 5% CO₂.