A Scalable Approach for the Generation of Human Pluripotent Stem Cell-Derived Hepatic Organoids with Sensitive Hepatotoxicity Features

• Published in: Stem cells and development Vol. 26; no. 20; p. 1490
• Main Authors: Sgodda, Malte, Dai, Zhen, Zweigerdt, Robert, Sharma, Amar Deep, Ott, Michael, Cantz, Tobias
• Format: Journal Article
• Language: English
• Published: United States 15.10.2017
• Subjects: Animals; Cell Culture Techniques - methods; Cell Differentiation; Cells, Cultured; Humans; Liver - pathology; Mice; Organoids - pathology; Pluripotent Stem Cells - cytology; organoid culture; toxicity assays; scalable culture; hepatic PSC derivatives
• ISSN: 1557-8534
• DOI: 10.1089/scd.2017.0023

The quest for physiologically active human hepatocyte-like cells for in vitro research and drug screening is high. The recent progress in the field of pluripotent stem cell (PSC)-derived hepatic cells within the last decade brings those cells closer to applications in translational medicine. However, the classical two-dimensional (2D) cell culture systems are of limited use, because relevant cell-cell interactions based on cell polarity, which is a major prerequisite for proper hepatic cell metabolisms, are not provided. In this study, we report a scalable 3D suspension culture system, in which PSC-derived hepatic cells can be maintained for up to 3 weeks with stable gene expression profiles and metabolic features in a suspension culture system ranging from a 1.5 mL up to a 15 mL. Adjustments of culture conditions and, most importantly, the size of the organoids resulted in the robust generation of hepatic organoids consisting of a quite homogenous cell population. Importantly, the generation of these hepatic organoids was highly reproducible and allowed, in contrast to hepatic PSC derivatives in 2D culture conditions, a sensitive assessment of acetaminophen-related toxicity, the most common source for drug-induced liver failure.