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Nat Protoc. 2015 Dec;10(12):2027-53. doi: 10.1038/nprot.2015.128. Epub 2015 Nov 19.

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

Author information

1
Broad Institute of MIT and Harvard, Cambridge, Massachusetts, USA.
2
Department of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
3
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
4
Laboratory of Virology and Infectious Disease, Center for the Study of Hepatitis C, The Rockefeller University, New York, New York, USA.
5
Unidade de Malaria, Instituto de Medicina Molecular, Universidade de Lisboa, Lisboa, Portugal.
6
Department of Mechanical Engineering, School of Biomedical Engineering, Colorado State University, Fort Collins, Colorado, USA.
7
Department of Medicine, Brigham and Women's Hospital, Boston, Massachusetts, USA.
8
Department of Electrical Engineering and Computer Science, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
9
Howard Hughes Medical Institute, Cambridge, Massachusetts, USA.
10
Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.

Abstract

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.

PMID:
26584444
PMCID:
PMC5867906
DOI:
10.1038/nprot.2015.128
[Indexed for MEDLINE]
Free PMC Article

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