Fig. 5.

Long-term culture of mature HepaRG cells in 3D printed following aflatoxin B1 treatment. (A) 3D constructs treated with AFB1 (10 μM or 20 μM) for one week then constructs were cultured for additional 2 weeks with the toxin showed reduction in cell viability as visualized using the live-dead staining; calcein-AM (living cells in green) and ethidium homodimer-1 (dead cells in red). (B) Number of green-stained cells quantified with ImageJ revealed a significant decrease in number of living cells after two and three weeks of culture. Bars indicate the means ± SD, n ≥ 3 images. (C) Metabolic activity of the mature HepaRG cells treated with DMSO or AFB1 (10 μM or 20 μM) inside the 3D printed alginate/gelatin/Matrigel constructs was determined by the XTT assay at the indicated time points. Absorbance was measured at 450 nm. (D) Albumin secretion from mature HepaRG cells cultured in the 3D constructs upon treatment with DMSO or AFB1 (10 μM or 20 μM) was quantified at the indicated time points using ELISA analysis. Results are shown as mean ± SD of three independent experiments. * p ≤ 0.05; ** p ≤ 0.01; *** p ≤ 0.001; **** p ≤ 0.0001.