Journal
NATURE PROTOCOLS
Volume 8, Issue 2, Pages 430-437Publisher
NATURE PUBLISHING GROUP
DOI: 10.1038/nprot.2012.153
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Funding
- Medical Research Council (MRC) Senior Non-clinical Fellowship
- Cambridge Hospitals National Institute for Health Research Biomedical Research Center
- EU
- Evelyn Trust
- Children's Liver Disease Foundation
- Medical Research Council [G0701448] Funding Source: researchfish
- MRC [G0701448] Funding Source: UKRI
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Large-scale production of hepatocytes from a variety of genetic backgrounds would be beneficial for drug screening and to provide a source of cells to be used as a substitute for liver transplantation. However, fully functional primary hepatocytes remain difficult to expand in vitro, and circumventing this problem by using an alternative source of cells is desirable. Here we describe a 25-d protocol to direct the differentiation of human pluripotent stem cells into a near-homogenous population of hepatocyte-like cells. As cells progress through this protocol, they express genes in a chronological manner similar to that described during in vivo hepatic development. The protocol relies on culture systems devoid of serum, feeders or complex extracellular matrices, which enable molecular analyses without interference from unknown factors. This approach works efficiently with human embryonic stem cells and human induced pluripotent stem cells and was recently used to model liver diseases in vitro.
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