A microfluidically perfused three dimensional human liver model

Knut Rennert; Sandra Steinborn; Marko Gröger; Birgit Ungerböck; Anne-Marie Jank; Josef Ehgartner; Sandor Nietzsche; Julia Dinger; Michael Kiehntopf; Harald Funke; Frank T Peters; Amelie Lupp; Claudia Gärtner; Torsten Mayr; Michael Bauer; Otmar Huber; Alexander S Mosig

doi:10.1016/j.biomaterials.2015.08.043

A microfluidically perfused three dimensional human liver model

Biomaterials. 2015 Dec:71:119-131. doi: 10.1016/j.biomaterials.2015.08.043. Epub 2015 Aug 25.

Authors

Knut Rennert¹, Sandra Steinborn¹, Marko Gröger², Birgit Ungerböck³, Anne-Marie Jank⁴, Josef Ehgartner⁵, Sandor Nietzsche⁶, Julia Dinger⁷, Michael Kiehntopf⁸, Harald Funke⁴, Frank T Peters⁷, Amelie Lupp⁹, Claudia Gärtner³, Torsten Mayr⁵, Michael Bauer¹⁰, Otmar Huber¹¹, Alexander S Mosig¹²

Affiliations

¹ Institute of Biochemistry II, Jena University Hospital, 07743 Jena, Germany.
² Institute of Biochemistry II, Jena University Hospital, 07743 Jena, Germany; Clinic of Anesthesiology and Intensive Care, Jena University Hospital, 07747 Jena, Germany.
³ Microfluidic ChipShop GmbH, Stockholmer Straße 20, 07747 Jena, Germany.
⁴ Molecular Hemostaseology, Jena University Hospital, Jena, 07743 Jena, Germany.
⁵ Institute of Analytical Chemistry and Food Chemistry, Graz University of Technology, Graz, Austria.
⁶ Center for Electron Microscopy, Jena University Hospital, 07743 Jena, Germany.
⁷ Institute of Forensic Medicine, University Hospital Jena, 07743 Jena, Germany.
⁸ Department of Clinical Chemistry and Laboratory Medicine, Jena University Hospital, 07743 Jena, Germany.
⁹ Department of Pharmacology and Toxicology, Jena University Hospital, 07747 Jena, Germany.
¹⁰ Clinic of Anesthesiology and Intensive Care, Jena University Hospital, 07747 Jena, Germany; Center for Sepsis Control and Care, Jena University Hospital, 07747 Jena, Germany.
¹¹ Institute of Biochemistry II, Jena University Hospital, 07743 Jena, Germany; Center for Sepsis Control and Care, Jena University Hospital, 07747 Jena, Germany.
¹² 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.

PMID: 26322723
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.

Keywords: Dynamic cell culture; Liver; Microfluidic biochip; Organoid; Oxygen.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Cells, Cultured
Humans
Liver / metabolism*
Microfluidics*
Microscopy, Electron, Scanning
Microscopy, Fluorescence
Multidrug Resistance-Associated Protein 2