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Additional file 1: Fig. S1. A lower degree of tumor differentiation correlates with poor prognosis in patients with pancreatic ductal adenocarcinoma (PDAC). Kaplan���Meier overall survival analysis based on histopathological tumor differentiation in patients with PDAC (n = 242). Fig. S2. The serum response of PDOs differed according to the tumor grade and Moffitt���s classification. Fig. S3. Spearman���s correlation analysis between the niche factor dependency and the expression levels of PDAC stem cell markers, including CD44, CD24, and CD133. Fig. S4. Supplementation of niche factors to Grade1 PDOs following indirect co-culture with CAFs allowed organoid formations. Fig. S5. The relationship between Moffitt���s classification (���Classical���and���Basal-like���) and the proliferation rate of PDOs in the absence of the indicated factors (**P

Additional file 2: Table S1. Relationship betweentumor differentiation and clinicopathologic factors.

Additional file 3: Supplementary Figure S1. Analyses of α-SMA and CK19 immunofluorescence staining in SUIT-2 or PSCs isolated from PDAC patients. PSCs presented a fibroblast-like appearance and expressed α-SMA, but not CK19 (original magnification: × 200). Pancreatic cancer cell (PCCs) line SUIT-2 expressed CK19, but not α-SMA. Supplementary Figure S2. Toxicity effects of NAC to PSCs and PCCs. (A-B) IC50 values were measured in PSC1–3 and PCCs by determined by CellTiter-Glo luminescent cell viability assay after 72 h treatment with indicated NAC concentrations. (C) Representative microscopy time-lapse images of PSC1 and PSC2 cells treated with 6 mM NAC in which concentration toxic to PSCs. (D) (Left) PSC1 and PSC2 cells were treated with 2.5 mM NAC for 48 h, stained with annexin V-FITC/PI, and then analyzed by flow cytometry. (Right) Quantification of apoptotic cells induced by NAC treatment. (E) the expression of cleaved caspase-3, a marker of apoptosis, was determined by western blotting in PSCs. PSCs were treated with PBS (as control), 2.5 mM NAC, or 20μM H2O2 (positive control). (F) the expression of α-SMA, an active marker of PSC, was determined by western blotting in PSC2, which treated with different concertation of NAC with or without supernatant from SUIT-2 (SUIT-SN). (G) Migration of PSC2 after 1 mM NAC treatment with or without SUIT-SN was analyzed. Migrated cell numbers were normalized by total cell numbers. H&E staining; original magnification, × 100. Scale bar = 100 μm. (H-I) Migration and invasion assays of PSCs and PCCs from Fig. 1c-d. Graphs show numbers of cells calculated from five fields. The exact number of migrated cells was counted. Scale bar = 100 μm. H&E staining; original magnification, × 100. (J) Effects of NAC were performed in PCCs and PSCs on mRNA expression of Nrf2, HMOX-1, NQO1, and GCLC. PBS was used as Control. *P