Journal Article

Micropatterned coculture of primary human hepatocytes and supportive cells for the study of hepatotropic pathogens

Trehan, Kartik, Gural, Nil, March, Sandra, Scull, Margaret A, Ramanan, Vyas, Mota, Maria M, Fleming, Heather E, Rice, Charles M, Bhatia, Sangeeta N, Galstian, Ani, Ng, Shengyong, Khetani, Salman R, Shlomai, Amir

2015

Overview

This protocol uses micropatterned co-cultures comprising 2D islands of primary human hepatocytes surrounded by supportive fibroblast cells to model liver infection by the hepatitis B and C viruses or Plasmodium pathogens in vitro . The development of therapies and vaccines for human hepatropic pathogens requires robust model systems that enable the study of host-pathogen interactions. However, in vitro liver models of infection typically use either hepatoma cell lines that exhibit aberrant physiology or primary human hepatocytes in culture conditions in which they rapidly lose their hepatic phenotype. To achieve stable and robust in vitro primary human hepatocyte models, we developed micropatterned cocultures (MPCCs), which consist of primary human hepatocytes organized into 2D islands that are surrounded by supportive fibroblast cells. By using this system, which can be established over a period of days, and maintained over multiple weeks, we demonstrate how to recapitulate in vitro hepatic life cycles for the hepatitis B and C viruses and the Plasmodium pathogens P. falciparum and P. vivax . The MPCC platform can be used to uncover aspects of host-pathogen interactions, and it has the potential to be used for drug and vaccine development.

• Publisher: Nature Publishing Group UK,Nature Publishing Group
• Subject: 631/1647/1407/651 / 631/1647/2234 / 631/326/417/2546 / 631/326/596/2553 / Analytical Chemistry / Animals / Biological Techniques / Cell culture / Cell Line / Cell lines / Cells, Cultured / Coculture Techniques - methods / Collagen / Computational Biology/Bioinformatics / Computer engineering / Drug resistance / Fibroblasts / Fibroblasts - cytology / Genotype & phenotype / Hepacivirus - physiology / Hepatitis / Hepatitis B / Hepatitis B - metabolism / Hepatitis B virus - physiology / Hepatitis C / Hepatitis C - metabolism / Hepatocytes / Hepatocytes - cytology / Hepatocytes - parasitology / Hepatocytes - virology / Hepatoma / Host-Pathogen Interactions / Humans / Infections / Islands / Life Sciences / Liver / Liver cancer / Liver cells / Malaria / Malaria - metabolism / Medical research / Methods / Mice / Microarrays / Micropatterning / Organic Chemistry / Pathogens / Phenotypes / Physiological aspects / Plasmodium / Plasmodium - physiology / Plasmodium falciparum - physiology / Plasmodium vivax - physiology / protocol / Quality control / Robustness / Tissue Array Analysis - methods / Vaccine development / Vaccines / Viruses