Hepatocellular carcinoma xenograft supports HCV replication: A mouse model for evaluating antivirals

Figure 1 Summary of overall experimental plan to generate a mouse-adapted hepatitis C virus-green fluorescence protein replicon cell line. A chimeric replicon clone was prepared by inserting the green fluorescence protein (GFP) coding sequences with the NS5A sequences of the hepatitis C virus (HCV) sub-genomic clone. Huh-7 cells were transfected with a transcribed sub-genomic HCV-RNA replicon. A stable Huh-7 cell line with replicating HCV GFP chimera RNA (S3-GFP replicon) was developed. The replicon cell line was implanted in SCID mice for tumor development. Subcutaneous tumor that developed in SCID mice was collected and cells with replicating HCV-GFP were selected by culture in growth medium that contained G-418 (1 mg/mL). The in vivo adaptation process was repeated to generate a mouse-adapted S3-GFP replicon cell line that demonstrated 50% GFP expression in subcutaneous tumors.