Abstract
Human induced pluripotent stem cells (h-iPSCs) represent a potentially unlimited source for the generation of human hepatocyte-like cells (h-iHLCs) for the establishment of platforms to study drug-induced hepatotoxicity, liver disease modeling, and ultimately the application of h-iHLCs in treatment of patients with end-stage liver disease. To understand the impact of donor-specific factors on the generation of h-iHLCs, the model for the direct comparison of h-iHLCs and primary human hepatocytes (PHHs) from the same human donor is needed. This study proposes a step-by-step protocol for plating, expansion, and characterization of primary human hepatic non-parenchymal cells (h-NPCs) isolated from the human liver, the reprogramming of generated h-NPCs into h-iPSCs and subsequent differentiation of reprogrammed h-iPSCs into h-iHLCs. The ultimate goal is to compare the gene expression involved in hepatocyte metabolism between h-iHLCs and PHHs from the same human donor thus eliminating interdonor variability. This newly developed protocol of h-NPC culture, expansion, and reprogramming into h-iPSCs allows: (1) utilization of a single organ source (i.e., liver) for isolation of PHHs and h-NPCs and (2) the in-depth study of donor factors involved in the generation of h-iHLCs with direct comparison to PHHs from the same donor.
Original language | English (US) |
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Article number | e112 |
Journal | Current Protocols in Stem Cell Biology |
Volume | 53 |
Issue number | 1 |
DOIs | |
State | Published - Jun 1 2020 |
Bibliographical note
Publisher Copyright:© 2020 Wiley Periodicals LLC
Keywords
- h-NPCs
- h-iPSCs
- human hepatic non-parenchymal cells
- human liver
- reprogramming