Scalable Spheroid Model of Human Hepatocytes for Hepatitis
C Infection and Replication
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Abstract
Developing effective new drugs against
hepatitis C (HCV) virus
has been challenging due to the lack of appropriate small animal and <i>in vitro</i> models recapitulating the entire life cycle of
the virus. Current <i>in vitro</i> models fail to recapitulate
the complexity of human liver physiology. Here we present a method
to study HCV infection and replication on spheroid cultures of Huh
7.5 cells and primary human hepatocytes. Spheroid cultures are constructed
using a galactosylated cellulosic sponge with homogeneous macroporosity,
enabling the formation and maintenance of uniformly sized spheroids.
This facilitates easy handling of the tissue-engineered constructs
and overcomes limitations inherent of traditional spheroid cultures.
Spheroids formed in the galactosylated cellulosic sponge show enhanced
hepatic functions in Huh 7.5 cells and maintain liver-specific functions
of primary human hepatocytes for 2 weeks in culture. Establishment
of apical and basolateral polarity along with the expression and localization
of all HCV specific entry proteins allow for a 9-fold increase in
viral entry in spheroid cultures over conventional monolayer cultures.
Huh 7.5 cells cultured in the galactosylated cellulosic sponge also
support replication of the HCV clone, JFH (Japanese fulminant hepatitis)-1
at higher levels than in monolayer cultures. The advantages of our
system in maintaining liver-specific functions and allowing HCV infection
together with its ease of handling make it suitable for the study
of HCV biology in basic research and pharmaceutical R&D