Microfluidic Hydrostatic Deposition Patterning for a confined hepatocyte-biliary epithelial cell co-culture system

Abstract

Reconstruction of hepato-biliary networks is very important to maintaining liver organoids in vitro, since the accumulation of bile has been proven to be toxic to hepatocytes, especially in long-term culture. We have developed culture methods for reconstructing bile canaliculi (BC) and bile ducts (BDs) formed by small hepatocytes (SHs) which are hepatic progenitor cells, and biliary epithelial cells (BEes), respectively. To study the mechanism of biliary excretion, we need to establish a co-culture system for SHs and BEes. A limitation of conventional culture methods in investigating the mechanisms of hepato-biliary connections is the difficulty to observe the dynamic interactions between SHs and BEes in culture. In addition, patterned constructs do not form in conventional cultures due to the lack of organization of cells. A system that allows more complex dynamics between SHs and BEes to occur is needed in order to investigate the interactions between these cell types. Here, we present an approach called Microfluidic Hydrostatic Deposition Patterning (MHDP) that patterns SH and BEe colonies with defined shape and position, which promotes productive interactions between the cells and eventually induce the formation of heterogeneous tissues integration which is thought to be a prototype of complete biliary networks.

Liver Tissue Engineering: Challenges and Opportunities

“…Lee et al. and Auth et al demonstrated the formation of the efficient hepato-biliary network under coculture conditions (hepatocytes and cholangiocytes). , In addition, a number of tissue-engineered artificial bile ducts have been reported. − However, integration of bile network and its incorporation in the 3D implantable constructs still remains less explored. In this regard, the whole liver decellularized matrix has gained importance because of retention of the intact biliary network .…”