Development of an Induced Pluripotent Stem Cell-Based Liver-on-a-Chip Assessed with an Alzheimer’s Disease Drug

Abstract

Liver-related drug metabolism is a key aspect of pharmacokinetics and possible toxicity. From this perspective, the availability of advanced in vitro models for drug testing is still an open need, also to the end of reducing the burden of in vivo experiments. In this scenario, organ-on-a-chip is gaining attention as it couples a state-of-the art in vitro approach to the recapitulation of key in vivo physiological features such as fluidodynamics and a tri-dimensional cytoarchitecture. We implemented a novel liver-on-a-chip (LoC) device based on an innovative dynamic device (MINERVA 2.0) where functional hepatocytes (iHep) have been encapsulated into a 3D hydrogel matrix interfaced through a porous membrane with endothelial cells (iEndo)]. Both lines were derived from human-induced pluripotent stem cells (iPSCs), and the LoC was functionally assessed with donepezil, a drug approved for Alzheimer’s disease therapy. The presence of iEndo and a 3D microenvironment enhanced the expression of liver-specific physiologic functions as in iHep, after 7 day perfusion, we noticed an increase of albumin, urea production, and cytochrome CYP3A4 expression compared to the iHep static culture. In particular, for donepezil kinetics, a computational fluid dynamic study conducted to assess the amount of donepezil diffused into the LoC indicated that the molecule should be able to pass through the iEndo and reach the target iHep construct. Then, we performed experiments of donepezil kinetics that confirmed the numerical simulations. Overall, our iPSC-based LoC reproduced the in vivo physiological microenvironment of the liver and was suitable for potential hepatotoxic screening studies.

Human gut epithelium features recapitulated in MINERVA 2.0 millifluidic organ-on-a-chip device

“…This is supported also by a comparison of this work with another manuscript of our group already published. 36 By comparing the two models, we can appreciate the versatility of MINERVA 2.0 in terms of type of cell culture (e.g., iPSC vs immortalized cells), different use of culture conditions based on the Transwell-like inserts (e.g., on both sides of the membrane, keeping the two fluid paths separated vs on one side of the membrane), cell seeding mode (e.g., cells embedded in a gel vs cells seeded directly on the membrane), and different organ development (e.g., liver vs intestine). 36 Moreover, the two glass inserts of MINERVA 2.0 in both the culture chambers allow optical access by light transmission or confocal microscopy, a desirable feature as already reported in other solutions.…”