A microfluidically perfused three dimensional human liver model

• Published in: Biomaterials Vol. 71; pp. 119 - 131
• Main Authors: Rennert, Knut, Steinborn, Sandra, Gröger, Marko, Ungerböck, Birgit, Jank, Anne-Marie, Ehgartner, Josef, Nietzsche, Sandor, Dinger, Julia, Kiehntopf, Michael, Funke, Harald, Peters, Frank T, Lupp, Amelie, Gärtner, Claudia, Mayr, Torsten, Bauer, Michael, Huber, Otmar, Mosig, Alexander S
• Format: Journal Article
• Language: English
• Published: Netherlands Elsevier Ltd 01.12.2015
• Subjects: Advanced Basic Science; Cells, Cultured; Cellular; Dentistry; Dynamic cell culture; Human; Humans; Liver; Liver - metabolism; Microfluidic biochip; Microfluidics; Microscopy, Electron, Scanning; Microscopy, Fluorescence; Nutrition; Organoid; Oxygen; Physiology; Secretions; Three dimensional; Transporter; Organoid; Microfluidic biochip; Oxygen; Liver; Dynamic cell culture
• ISSN: 0142-9612; 1878-5905
• DOI: 10.1016/j.biomaterials.2015.08.043

Abstract Within the liver, non-parenchymal cells (NPCs) are critically involved in the regulation of hepatocyte polarization and maintenance of metabolic function. We here report the establishment of a liver organoid that integrates NPCs in a vascular layer composed of endothelial cells and tissue macrophages and a hepatic layer comprising stellate cells co-cultured with hepatocytes. The three-dimensional liver organoid is embedded in a microfluidically perfused biochip that enables sufficient nutrition supply and resembles morphological aspects of the human liver sinusoid. It utilizes a suspended membrane as a cell substrate mimicking the space of Disse. Luminescence-based sensor spots were integrated into the chip to allow online measurement of cellular oxygen consumption. Application of microfluidic flow induces defined expression of ZO-1, transferrin, ASGPR-1 along with an increased expression of MRP-2 transporter protein within the liver organoids. Moreover, perfusion was accompanied by an increased hepatobiliary secretion of 5(6)-carboxy-2′,7′-dichlorofluorescein and an enhanced formation of hepatocyte microvilli. From this we conclude that the perfused liver organoid shares relevant morphological and functional characteristics with the human liver and represents a new in vitro research tool to study human hepatocellular physiology at the cellular level under conditions close to the physiological situation.