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Human Liver Sinusoid on a Chip for Hepatitis B Virus Replication Study

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 con...

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Autores principales: Kang, Young Bok (Abraham), Rawat, Siddhartha, Duchemin, Nicholas, Bouchard, Michael, Noh, Moses
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6190364/
http://dx.doi.org/10.3390/mi8010027
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author Kang, Young Bok (Abraham)
Rawat, Siddhartha
Duchemin, Nicholas
Bouchard, Michael
Noh, Moses
author_facet Kang, Young Bok (Abraham)
Rawat, Siddhartha
Duchemin, Nicholas
Bouchard, Michael
Noh, Moses
author_sort Kang, Young Bok (Abraham)
collection PubMed
description 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.
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spelling pubmed-61903642018-11-01 Human Liver Sinusoid on a Chip for Hepatitis B Virus Replication Study Kang, Young Bok (Abraham) Rawat, Siddhartha Duchemin, Nicholas Bouchard, Michael Noh, Moses Micromachines (Basel) Article 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. MDPI 2017-01-20 /pmc/articles/PMC6190364/ http://dx.doi.org/10.3390/mi8010027 Text en © 2017 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Kang, Young Bok (Abraham)
Rawat, Siddhartha
Duchemin, Nicholas
Bouchard, Michael
Noh, Moses
Human Liver Sinusoid on a Chip for Hepatitis B Virus Replication Study
title Human Liver Sinusoid on a Chip for Hepatitis B Virus Replication Study
title_full Human Liver Sinusoid on a Chip for Hepatitis B Virus Replication Study
title_fullStr Human Liver Sinusoid on a Chip for Hepatitis B Virus Replication Study
title_full_unstemmed Human Liver Sinusoid on a Chip for Hepatitis B Virus Replication Study
title_short Human Liver Sinusoid on a Chip for Hepatitis B Virus Replication Study
title_sort human liver sinusoid on a chip for hepatitis b virus replication study
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6190364/
http://dx.doi.org/10.3390/mi8010027
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