Abstract
We have developed a miniature human liver (liver-sinusoid-on-a-chip) model using a dual microchannel separated by a porous membrane. Primary human hepatocytes and immortalized bovine aortic endothelial cells were co-cultured on opposite sides of a microporous membrane in a dual microchannel with continuous perfusion. Primary human hepatocytes in this system retained their polygonal morphology for up to 26 days, while hepatocytes cultured in the absence of bovine aortic endothelial cells lost their morphology within a week. In order to demonstrate the utility of our human-liver-sinusoid-on-a-chip, human hepatocytes in this system were directly infected by Hepatitis B Virus (HBV). Expression of the HBV core antigen was detected in human hepatocytes in the microchannel system. HBV replication, measured by the presence of cell-secreted HBV DNA, was also detected. Importantly, HBV is hepatotropic, and expression of HBV RNA transcripts is dependent upon expression of hepatocyte-specific factors. Moreover, HBV infection requires expression of the human-hepatocyte-specific HBV cell surface receptor. Therefore, the ability to detect HBV replication and Hepatitis B core Antigen (HBcAg) expression in our microfluidic platform confirmed that hepatocyte differentiation and functions were retained throughout the time course of our studies. We believe that our human-liver-sinusoid-on-a-chip could have many applications in liver-related research and drug development.
Highlights
The World Health Organization has estimated that approximately 240 million people in the world have a chronic Hepatitis B Virus (HBV) infection of the liver [1,2]
Hepatocytes are naturally infected with HBV that is introduced into the liver sinusoid via the blood stream (Figure 1); because our goal was to demonstrate that hepatocytes cultured in our system can be directly infected by HBV, we directly introduced HBV into the bottom channel, where hepatocytes are cultured in order to improve the efficiency of viral infection
When primary human hepatocytes-only were cultured on a microporous membrane of a dual-PDMS microchannel under continuous perfusion, these hepatocytes maintained a normal phenotype in the microchannel for about 5–7 days after seeding, which was longer than hepatocytes that were cultured alone under static conditions (Figure 2c,d)
Summary
The World Health Organization has estimated that approximately 240 million people in the world have a chronic Hepatitis B Virus (HBV) infection of the liver [1,2]. Micromachines 2017, 8, 27 for this gap in our knowledge is the absence of in vitro human hepatocyte culture systems that mimic the environment and cellular architecture of the liver and can be used for long-term studies of the consequence of an HBV infection to human hepatocyte physiology. The development of these types of systems is critical for providing platforms that can be used to define HBV effects on hepatocyte physiology.
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
