Center for the Study of Hepatitis C and Laboratory of Virology and Infectious Diseases, The Rockefeller University, New York, NY 10065, USA.
Proc Natl Acad Sci U S A. 2010 Feb 16;107(7):3141-5. doi: 10.1073/pnas.0915130107. Epub 2010 Feb 1.
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.
丙型肝炎病毒 (HCV) 仍然是一个主要的公共卫生问题，影响着全球约 1.3 亿人。HCV 感染可导致肝硬化、肝细胞癌和终末期肝病，以及肝外并发症，如冷球蛋白血症和淋巴瘤。预防和治疗的选择非常有限；目前尚无 HCV 疫苗，非特异性 IFN 治疗通常无效。由于缺乏可靠的 HCV 细胞培养系统来准确预测人体反应，靶向抗病毒药物的开发受到阻碍。在这里，我们展示了在多井格式的原代人肝细胞和支持基质的微图案共培养物 (MPCCs) 中重现的整个 HCV 生命周期。MPCCs 形成极化细胞层，表达所有已知的 HCV 进入因子，并持续复制数周。当与高灵敏度的荧光和发光报告系统结合使用时，MPCCs 有可能成为一种高通量平台，用于同时评估抗 HCV 治疗药物的体外疗效和毒性特征。
