Z Gastroenterol 2012; 50 - P2_03
DOI: 10.1055/s-0031-1295800

A Microfluidic 3D Hepatocyte Culture System -HepaChip®- For Drug And Toxicity Investigations

J Böttger 1, J Schütte 2, K Benz 2, C Freudigmann 2, B Hagmeyer 2, C Höppner 3, H Becker 4, P Röhnert 5, M Stelzle 2, R Gebhardt 1
  • 1Institut für Biochemie, Medizinische Fakultät, Universität Leipzig, Leipzig, Germany
  • 2NMI Naturwissenschaftliches und Medizinisches Institut an der Universität Tübingen, Reutlingen
  • 3BSL BIOSERVICE Scientific Laboratories GmbH, Planegg
  • 4microfluidic ChipShop GmbH, Jena
  • 5European Screeningport GmbH, Hamburg

PRESENTLY used in vitro test systems for drug and toxicity investigations lack the complexity of the liver and reduce test systems solely to hepatocytes or random co-cultures with non-parenchymal cells. Unfortunately, hepatocytes rapidly lose their typical morphology and the expression of metabolizing enzymes, which leads to cell phenotypes that differ from those found in the human physiology. This unstable phenotype is one of the main reasons for the failure to correctly predict drug-induced liver toxicity.

TO ADDRESS THIS, we are developing a microfluidic HepaChip® culture system, which mimics the liver acinus architecture to restore the 3D structure and function of primary hepatocytes, maintain their differentiation status and achieve longer incubation periods. Furthermore, the flow conditions in HepaChip® cultures facilitate the physiological-like liver zonation and shear stress. HepaChip® comprises microstructured ridges (50×500µm) which are bio-functionalized with collagen IV to provide cells with extracellular matrix. 3D artificial sinusoids are obtained by assembling primary or cryopreserved human and mice hepatocytes on bio-functionalized areas using dielectrophoretic and hydrodynamic forces. Subsequently, human liver endothelial cells are assembled in the same manner forming a second cell layer on the side and top and of hepatocytes to establish sinusoid-like 3D cords.

RESULTS: Due to perfusion, the artificial hepatocyte cords exhibit a certain stable phenotype over several days while morphological alterations were observed solely in ordinary 2D cultures. This is accompanied by a remarkable increase in metabolic performance for phase I, phase II, CYP1A2 and 3A4 enzyme activity in HepaChip®. Under these in vivo-like conditions the formation of proper cell-cell interactions and polarized hepatocytes is possible, which improves drug investigations by increasing the predictability of biological and metabolic effects to xenobiotics and natural products.

3D hepatocyte culture - cell culture chip - metabolism - microfluidic - toxicity