Spheroid models of liver cells for Hepatitis C infections

摘要

With over 170 million people affected worldwide, Hepatitis C is a major health problem in the developed and developing world. The lack of preventative therapeutic options and ineffective drugs to cure or curtail the progression of the disease, subjects health systems around the world to huge burdens. Hepatitis C virus primarily replicates only in the human liver and the lack of appropriate tissue engineered models of the liver cells which maintain the differentiated phenotype of these cells has greatly compromised the development of novel therapeutic compounds. Here we demonstrate the utility of a novel macroporous galactosylated cellulosic scaffold with mechanical properties which closely resembles the native liver to form spheroids of Huh 7.5 cells a model cell line for HCV infections and primary human hepatocytes. The well controlled macroporosity of the scaffold facilitates the formation of spheroids of uniform size and distribution. The tissue engineered models of both these cell types form polarized spheroids with appropriate expression and localisation of viral entry receptors namely CD-81, Claudin-1, Occludin and SCARB-1. The spheroid cultures being more physiologically relevant also elevate the differentiated phenotype of Huh 7.5 cells and maintain the differentiated phenotype for primary human hepatocytes over 14 days in culture as determined by qRT-PCR. These results affirm the utility of the spheroid model to study HCV infection and life cycle. We will further test the system for HCV infection using pseudoparticles and the live Hepatitis C virus.