title: Liver sinusoid on a chip: Long-term layered co-culture of primary rat hepatocytes and endothelial cells in microfluidic platforms.
abstract: We describe the generation of microfluidic platforms for the co-culture of primary hepatocytes and endothelial cells; these platforms mimic the architecture of a liver sinusoid. This paper describes a progressional study of creating such a liver sinusoid on a chip system. Primary rat hepatocytes (PRHs) were co-cultured with primary or established endothelial cells in layers in single and dual microchannel configurations with or without continuous perfusion. Cell viability and maintenance of hepatocyte functions were monitored and compared for diverse experimental conditions. When primary rat hepatocytes were co-cultured with immortalized bovine aortic endothelial cells (BAECs) in a dual microchannel with continuous perfusion, hepatocytes maintained their normal morphology and continued to produce urea for at least 30 days. In order to demonstrate the utility of our microfluidic liver sinusoid platform, we also performed an analysis of viral replication for the hepatotropic hepatitis B virus (HBV). HBV replication, as measured by the presence of cell-secreted HBV DNA, was successfully detected. We believe that our liver model closely mimics the in vivo liver sinusoid and supports long-term primary liver cell culture. This liver model could be extended to diverse liver biology studies and liver-related disease research such as drug induced liver toxicology, cancer research, and analysis of pathological effects and replication strategies of various hepatotropic infectious agents. .
authors.affiliation: Mechanical Engineering and Mechanics, Drexel University, Philadelphia, Pennsylvania.
authors.forename: Young Bok Abraham
authors.initials: YB
authors.lastname: Kang
authors.affiliation: Mechanical Engineering and Mechanics, Drexel University, Philadelphia, Pennsylvania.
authors.forename: Temitope R
authors.initials: TR
authors.lastname: Sodunke
authors.affiliation: Graduate Program in Microbiology and Immunology, Drexel University College of Medicine, Philadelphia, Pennsylvania.
authors.forename: Jason
authors.initials: J
authors.lastname: Lamontagne
authors.affiliation: Mechanical Engineering and Mechanics, Drexel University, Philadelphia, Pennsylvania.
authors.forename: Joseph
authors.initials: J
authors.lastname: Cirillo
authors.affiliation: Graduate Program in Biochemistry, Drexel University College of Medicine, Philadelphia, Pennsylvania.
authors.forename: Caroline
authors.initials: C
authors.lastname: Rajiv
authors.affiliation: Department of Biochemistry and Molecular Biology, Drexel University College of Medicine, Philadelphia, Pennsylvania. michael.bouchard@drexelmed.edu.
authors.forename: Michael J
authors.initials: MJ
authors.lastname: Bouchard
authors.affiliation: Mechanical Engineering and Mechanics, Drexel University, Philadelphia, Pennsylvania. mosesnoh@coe.drexel.edu.
authors.forename: Moses
authors.initials: M
authors.lastname: Noh
mesh_terms: D000818:Animals; D002417:Cattle; D002470:Cell Survival; D002478:Cells, Cultured; D018920:Coculture Techniques; D004279:DNA, Viral / Q000032:analysis; D042783:Endothelial Cells / Q000502:physiology*; D006515:Hepatitis B virus / Q000502:physiology; D022781:Hepatocytes / Q000502:physiology*; D019164:Liver, Artificial; D044085:Microfluidics / Q000379:methods*; D051381:Rats; D013997:Time Factors; D014508:Urea / Q000378:metabolism; D014779:Virus Replication
chemical_list: D004279:DNA, Viral; D014508:Urea
keywords: liver model; liver on a chip; liver sinusoid; long-term co-culture; microfluidic platform