FIG. 3.

(a) Optimization of hepatocytes seeding density and culture medium. (i) Morphological and viability comparison between 20 and 40 M/ml seeding densities with a medium supplemented with ACA or without it. In the absence of ACA, the hepatocytes remodel the gel by impairing the circularity of the central lumen. Scale bars are 140 μm. (ii) Hepatocytes secrete higher albumin levels when seeded at 40 M/ml while (iii) CYP3A4 activity is similar between the two seeding densities. Statistical analysis was performed using analysis of variance (ANOVA) with the Bonferroni correction: ** p = 0.002, *** p < 0.001 (N = 3). (b) Comparison between static (DC-S) and dynamic (DC-D) conditions applied to the platform. (i) Pictures on day 1 (24 h after hepatocytes seeding) and day 9 (end of the culture). Hepatocytes maintain their characteristic morphology throughout the culture without any observable differences between the conditions. Scale bars are 140 μm. (ii) LDH release decreased from day 3 indicating high cellular viability. (iii) Hepatocytes produce higher levels of albumin in the dynamic condition (DC-D) compared to static condition (DC-S). (iv) CYP3A4 levels are similar between the two conditions.