Abstract
Background & aimsHepatocyte-like cells (HLCs) differentiated from induced pluripotent stem cells (iPSCs) have emerged as a promising cell culture model to study metabolism, biotransformation, viral infections and inherited liver diseases. iPSCs provide an unlimited supply for the generation of HLCs, but incomplete HLC differentiation remains a major challenge. iPSC may carry-on a tissue of origin dependent expression memory influencing iPSC differentiation into different cell types. Whether liver derived iPSCs (Li-iPSCs) would allow the generation of more fully differentiated HLCs is not known.MethodsIn the current study, we used primary liver cells (PLCs) expanded from liver needle biopsies and reprogrammed them into Li-iPSCs using a non-integrative Sendai virus-based system. Li-iPSCs were differentiated into HLCs using established differentiation protocols. The HLC phenotype was characterized at the protein, functional and transcriptional level. RNA sequencing data were generated from the originating liver biopsies, the Li-iPSCs, fibroblast derived iPSCs, and differentiated HLCs, and used to characterize and compare their transcriptome profiles.ResultsLi-iPSCs indeed retain a liver specific transcriptional footprint. Li-iPSCs can be propagated to provide an unlimited supply of cells for differentiation into Li-HLCs. Similar to HLCs derived from fibroblasts, Li-HLCs could not be fully differentiated into hepatocytes. Relative to the originating liver, Li-HLCs showed lower expression of liver specific transcription factors and increased expression of genes involved in the differentiation of other tissues.ConclusionsPLCs and Li-iPSCs obtained from small pieces of human needle liver biopsies constitute a novel unlimited source for the production of HLCs. Despite the preservation of a liver specific gene expression footprint in Li-iPSCs, the generation of fully differentiated hepatocytes cannot be achieved with the current differentiation protocols.
Highlights
Current in vitro models for the study of liver functions and pathology predominantly rely on hepatoma cell lines and primary human hepatocyte (PHH) cultures
Hepatocyte-like cells (HLCs) differentiated from induced pluripotent stem cells have emerged as a promising cell culture model to study metabolism, biotransformation, viral infections and inherited liver diseases. iPSCs provide an unlimited supply for the generation of hepatocyte-like cells (HLCs), but incomplete HLC differentiation remains a major challenge. iPSC may carry-on a tissue of origin dependent expression memory influencing iPSC differentiation into different cell types
Similar to HLCs derived from fibroblasts, LiiPSCs into hepatocyte-like cells (Li-HLCs) could not be fully differentiated into hepatocytes
Summary
Current in vitro models for the study of liver functions and pathology predominantly rely on hepatoma cell lines and primary human hepatocyte (PHH) cultures. Hepatoma cell lines are characterized by a stable phenotype, they show limited expression of liver enzymes [1] Since these cells are clonal in origin, they poorly represent the intra- and inter-patient cell heterogeneity. PHHs are considered the gold standard for toxicology studies because they maintain numerous hepatocyte functions Their hepatic phenotype is unstable over long term culturing, and their availability is limited (reviewed in [2]). Hepatocyte-like cells (HLCs) differentiated from induced pluripotent stem cells (iPSCs) have emerged as a promising cell culture model to study metabolism, biotransformation, viral infections and inherited liver diseases. Whether liver derived iPSCs (Li-iPSCs) would allow the generation of more fully differentiated HLCs is not known
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