"Artificial micro organs"--a microfluidic device for dielectrophoretic assembly of liver sinusoids.

In order to study possible toxic side effects of potential drug compounds in vitro a reliable test system is needed. Predicting liver toxicity presents a major challenge of particular importance as liver cells grown in a cell culture suffer from a rapid loss of their liver specific functions. Therefore we are developing a new microfluidic test system for liver toxicity. This test system is based on an organ-like liver 3D co-culture of hepatocytes and endothelial cells. We devised a microfluidic chip featuring cell culture chambers with integrated electrodes for the assembly of liver sinusoids by dielectrophoresis. Fluid channels enable an organ-like perfusion with culture media and test compounds. Different chamber designs were studied and optimized with regard to dielectrophoretic force distribution, hydrodynamic flow profile, and cell trapping rate using numeric simulations. Based on simulation results a microchip was injection-moulded from COP. This chip allowed the assembly of viable hepatocytes and endothelial cells in a sinusoid-like fashion.

Numerical modelling and measurement of cell trajectories in 3-D under the influence of dielectrophoretic and hydrodynamic forces.

This research is part of a program aiming at the development of a fluidic microsystem for in vitro drug testing. For this purpose, primary cells need to be assembled to form cellular aggregates in such a way as to resemble the basic functional units of organs. By providing for in vivo-like cellular contacts, proper extracellular matrix interaction and medium perfusion it is expected that cells will retain their phenotype over prolonged periods of time. In this way, in vitro test systems exhibiting in vivo type predictivity in drug testing are envisioned. Towards this goal a 3-D microstructure micro-milled in a cyclic olefin copolymer (COC) was designed in such a way as to assemble liver cells via insulator-based dielectrophoresis (iDEP) in a sinusoid-type fashion. First, numeric modelling and simulation of dielectrophoretic and hydrodynamic forces acting on cells in this microsystem was performed. In particular, the problem of the discontinuity of the electric field at the interface between the fluid media in the system and the polymer materials it consists of was addressed. It was shown that in certain cases, the material of the microsystem may be neglected altogether without introducing considerable error into the numerical solution. This simplification enabled the simulation of 3-D cell trajectories in complex chip geometries. Secondly, the assembly of HepG2 cells by insulator-based dielectrophoresis in this device is demonstrated. Finally, theoretical results were validated by recording 3-D cell trajectories and the Clausius-Mossotti factor of liver cells was determined by combining results obtained from both simulation and experiment.