Abstract
BackgroundCell therapy using hepatocytes derived from stem cells has been regarded as a promising alternate to liver transplantation. However, the heterogeneity of these hepatocytes makes them unsuitable for therapeutic use. To overcome this limitation, we generated homogenous hepatocyte like induced hepatocyte-like (iHep) cells.MethodsiHep cells were generated from induced pluripotent stem cells (iPSCs) integrated with the albumin (ALB) reporter gene. The therapeutic properties of these iHep cells were investigated after transplantation in fibrotic liver tissues of a mouse model.ResultsThe iHep cells expressed hepatocyte specific genes and proteins, and exhibited high levels of hepatocyte growth factor (HGF) and interleukin (IL)-10 expressions. Transplantation of iHep cells significantly decreased thioacetamide (TAA)-induced liver fibrosis, apoptotic cells in the liver, and ameliorated abnormal liver function. Liver tissues engrafted with iHep cells exhibited decreased expression of pro-inflammatory factors such as transforming growth factor (TGF)-β, IL-6, and monocyte chemo attractant protein (MCP)-1. Furthermore, an increased number of proliferating hepatocytes and human albumin-expressing iHep cells were detected in mice liver.ConclusionsThis study has investigated and proven the liver regeneration potential of genome-edited iHep cells and promises to be a strong foundation for further studies exploring cell therapy as an alternative therapeutic option for the treatment of liver fibrosis.
Highlights
Recent studies have demonstrated that functional hepatic lineage cells can be obtained from stem cells orChoi et al Stem Cell Research & Therapy (2020) 11:332 reporter genes is an effective way for separating differentiated cells from heterogeneous cell cultures.Lineage-specific reporter cells lines derived from embryonic stem cells (ES) or induced pluripotent stem cells can provide optimized strategy for the direct cell differentiation protocols
This study has investigated and proven the liver regeneration potential of genome-edited induced hepatocyte-like cells (iHep) cells and promises to be a strong foundation for further studies exploring cell therapy as an alternative therapeutic option for the treatment of liver fibrosis
To confirm the reporter system is functional, we measured the expression level of ALB protein in iHep using Western blot. iHep cells highly expressed ALB protein compare with the induced pluripotent stem cells (iPS) or uninduced iHep cells (uHep) (Supplemetary Fig. 3)
Summary
Recent studies have demonstrated that functional hepatic lineage cells can be obtained from stem cells orChoi et al Stem Cell Research & Therapy (2020) 11:332 reporter genes is an effective way for separating differentiated cells from heterogeneous cell cultures.Lineage-specific reporter cells lines derived from embryonic stem cells (ES) or induced pluripotent stem cells (iPS) can provide optimized strategy for the direct cell differentiation protocols. Knock-in reporter cell lines have been generated using transcription factors, lineage specific genes and gene editing [10,11,12]. Hepatocyte cell lines, rat primary hepatocyte, bone marrow cells were transduced using adenoviral vector containing ALB promoter ZsGreen reporter gene [13]. Since adenoviruses do not integrate into host genomes, their use for gene transfer resulted in transient expression of the reporter system. This limited the longterm observation of the differentiated cells. The heterogeneity of these hepatocytes makes them unsuitable for therapeutic use To overcome this limitation, we generated homogenous hepatocyte like induced hepatocyte-like (iHep) cells
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