title: A microfluidically perfused three dimensional human liver model.
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.
authors: Institute of Biochemistry II, Jena University Hospital, 07743 Jena, Germany.
authors: Knut
authors: K
authors: Rennert
authors: Institute of Biochemistry II, Jena University Hospital, 07743 Jena, Germany.
authors: Sandra
authors: S
authors: Steinborn
authors: Institute of Biochemistry II, Jena University Hospital, 07743 Jena, Germany; Clinic of Anesthesiology and Intensive Care, Jena University Hospital, 07747 Jena, Germany.
authors: Marko
authors: M
authors: Gröger
authors: Microfluidic ChipShop GmbH, Stockholmer Straße 20, 07747 Jena, Germany.
authors: Birgit
authors: B
authors: Ungerböck
authors: Molecular Hemostaseology, Jena University Hospital, Jena, 07743 Jena, Germany.
authors: Anne-Marie
authors: AM
authors: Jank
authors: Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Graz, Austria.
authors: Josef
authors: J
authors: Ehgartner
authors: Center for Electron Microscopy, Jena University Hospital, 07743 Jena, Germany.
authors: Sandor
authors: S
authors: Nietzsche
authors: Institute of Forensic Medicine, University Hospital Jena, 07743 Jena, Germany.
authors: Julia
authors: J
authors: Dinger
authors: Department of Clinical Chemistry and Laboratory Medicine, Jena University Hospital, 07743 Jena, Germany.
authors: Michael
authors: M
authors: Kiehntopf
authors: Molecular Hemostaseology, Jena University Hospital, Jena, 07743 Jena, Germany.
authors: Harald
authors: H
authors: Funke
authors: Institute of Forensic Medicine, University Hospital Jena, 07743 Jena, Germany.
authors: Frank T
authors: FT
authors: Peters
authors: Department of Pharmacology and Toxicology, Jena University Hospital, 07747 Jena, Germany.
authors: Amelie
authors: A
authors: Lupp
authors: Microfluidic ChipShop GmbH, Stockholmer Straße 20, 07747 Jena, Germany.
authors: Claudia
authors: C
authors: Gärtner
authors: Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Graz, Austria.
authors: Torsten
authors: T
authors: Mayr
authors: Clinic of Anesthesiology and Intensive Care, Jena University Hospital, 07747 Jena, Germany; Center for Sepsis Control and Care, Jena University Hospital, 07747 Jena, Germany.
authors: Michael
authors: M
authors: Bauer
authors: Institute of Biochemistry II, Jena University Hospital, 07743 Jena, Germany; Center for Sepsis Control and Care, Jena University Hospital, 07747 Jena, Germany.
authors: Otmar
authors: O
authors: Huber
authors: Institute of Biochemistry II, Jena University Hospital, 07743 Jena, Germany; Center for Sepsis Control and Care, Jena University Hospital, 07747 Jena, Germany. Electronic address: alexander.mosig@med.uni-jena.de.
authors: Alexander S
authors: AS
authors: Mosig
mesh_terms: D002478:Cells, Cultured; D006801:Humans; D008099:Liver / Q000378:metabolism*; D044085:Microfluidics; D008855:Microscopy, Electron, Scanning; D008856:Microscopy, Fluorescence; D000089762:Multidrug Resistance-Associated Protein 2
keywords: Dynamic cell culture; Liver; Microfluidic biochip; Organoid; Oxygen