Integration of an oxygen sensor into a polydymethylsiloxane hepatic culture device for two-dimensional gradient characterization

Abstract

This paper presents a new hepatic culture device, in which oxygen concentration gradient formed by cellular respiration can be visualized as a two-dimensional optical image. An oxygen sensor layer containing Pt (II) octaethylporphyrin (PtOEP) was embedded at the bottom of the device, allowing visualization of oxygen concentration distribution over the cell culture area. Oxygen concentration was determined by analyzing the modulation of fluorescence intensity emitted by the sensor, which is negatively correlated to the concentration. The results in HepG2 cell culture showed that the oxygen gradient and local oxygen partial pressure could be regulated by controlling the perfusion flow rate in the device. The local and physiological oxygen gradient comparable to the in vivo liver lobules can successfully be reproduced in the device. Therefore, periportal and perivenous hepatic like-zones were successfully realized in vitro. The present device would be helpful in the development of novel in vitro methods recapitulating the liver physiology for investigation of hepatic zonation and drug screening system recapitulating the liver physiology.