Persistent hepatitis C virus infection in microscale primary human hepatocyte cultures.
Hepatitis C virus (HCV) remains a major public health problem, affecting approximately 130 million people worldwide. HCV infection can lead to cirrhosis, hepatocellular carcinoma, and end-stage liver disease, as well as extrahepatic complications such as cryoglobulinemia and lymphoma. Preventative and therapeutic options are severely limited; there is no HCV vaccine available, and nonspecific, IFN-based treatments are frequently ineffective. Development of targeted antivirals has been hampered by the lack of robust HCV cell culture systems that reliably predict human responses. Here, we show the entire HCV life cycle recapitulated in micropatterned cocultures (MPCCs) of primary human hepatocytes and supportive stroma in a multiwell format. MPCCs form polarized cell layers expressing all known HCV entry factors and sustain viral replication for several weeks. When coupled with highly sensitive fluorescence- and luminescence-based reporter systems, MPCCs have potential as a high-throughput platform for simultaneous assessment of in vitro efficacy and toxicity profiles of anti-HCV therapeutics.
Ploss
Alexander
A
Center for the Study of Hepatitis C and Laboratory of Virology and Infectious Diseases, The Rockefeller University, New York, NY 10065, USA.
Khetani
Salman R
SR
Jones
Christopher T
CT
Syder
Andrew J
AJ
Trehan
Kartik
K
Gaysinskaya
Valeriya A
VA
Mu
Kathy
K
Ritola
Kimberly
K
Rice
Charles M
CM
Bhatia
Sangeeta N
SN
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods*; D002478:Cells, Cultured; D016174:Hepacivirus / Q000502:physiology*; D006526:Hepatitis C / Q000503:physiopathology*; D022781:Hepatocytes / Q000821:virology*; D006801:Humans; D023822:Tissue Engineering / Q000379:methods*; D053586:Virus Internalization / Q000187:drug effects; D014779:Virus Replication / Q000502:physiology
D000911:Antibodies, Monoclonal