A 3D microfluidic liver model for high throughput compound toxicity screening in the OrganoPlate

• Published in: Toxicology (Amsterdam) 2021-02, Vol.450, p.152667, Article 152667
• Main Authors: Bircsak, Kristin M., DeBiasio, Richard, Miedel, Mark, Alsebahi, Alaa, Reddinger, Ryan, Saleh, Anthony, Shun, Tongying, Vernetti, Lawrence A., Gough, Albert
• Format: Article
• Language: English
• Subjects: Cell Culture Techniques - methods; Cell Survival - drug effects; Cell Survival - physiology; Cytochrome P-450 CYP3A - metabolism; Dose-Response Relationship, Drug; Hepatocytes - drug effects; Hepatocytes - physiology; Hepatotoxicity; High throughput; High-Throughput Screening Assays - methods; Humans; Induced Pluripotent Stem Cells - drug effects; Induced Pluripotent Stem Cells - physiology; iPSC-derived hepatocytes; Liver - cytology; Liver - drug effects; Liver - physiology; Liver-on-a-chip; Microfluidics - methods; Screen; Toxicity Tests - methods; Troglitazone; Troglitazone - toxicity
• ISSN: 0300-483X; 1879-3185
• DOI: 10.1016/j.tox.2020.152667

We report the development, automation and validation of a 3D, microfluidic liver-on-a-chip for high throughput hepatotoxicity screening, the OrganoPlate LiverTox™. The model is comprised of aggregates of induced pluripotent stem cell (iPSC)-derived hepatocytes (iHep) seeded in an extracellular matrix in the organ channel and co-cultured with endothelial cells and THP-1 monoblasts differentiated to macrophages seeded in the vascular channel of the 96 well Mimetas OrganoPlate 2-lane. A key component of high throughput screening is automation and we report a protocol to seed, dose, collect and replenish media and add assay reagents in the OrganoPlate 2-lane using a standard laboratory liquid handling robot. A combination of secretome measurements and image-based analysis was used to demonstrate stable 15 day cell viability, albumin and urea secretion. Over the same time-period, CYP3A4 activity increased and alpha-fetoprotein secretion decreased suggesting further maturation of the iHeps. Troglitazone, a clinical hepatotoxin, was chosen as a control compound for validation studies. Albumin, urea, hepatocyte nuclear size and viability staining provided Robust Z’factors > 0.2 in plates treated 72 h with 180 μM troglitazone compared with a vehicle control. The viability assay provided the most robust statistic for a Robust Z’ factor = 0.6. A small library of 159 compounds with known liver effects was added to the OrganoPlate LiverTox model for 72 h at 50 μM and the Toxicological Prioritization scores were calculated. A follow up dose-response evaluation of select hits revealed the albumin assay to be the most sensitive in calculating TC50 values. This platform provides a robust, novel model which can be used for high throughput hepatotoxicity screening.