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In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration
Open accessAbstract
The Wnt target gene Lgr5 (leucine-rich-repeat-containing G-protein-coupled receptor 5) marks actively dividing stem cells in Wnt-driven, self-renewing tissues such as small intestine and colon, stomach and hair follicles. A three-dimensional culture system allows long-term clonal expansion of single Lgr5(+) stem cells into transplantable organoids (budding cysts) that retain many characteristics of the original epithelial architecture. A crucial component of the culture medium is the Wnt agonist RSPO1, the recently discovered ligand of LGR5. Here we show that Lgr5-lacZ is not expressed in healthy adult liver, however, small Lgr5-LacZ(+) cells appear near bile ducts upon damage, coinciding with robust activation of Wnt signalling. As shown by mouse lineage tracing using a new Lgr5-IRES-creERT2 knock-in allele, damage-induced Lgr5(+) cells generate hepatocytes and bile ducts in vivo. Single Lgr5(+) cells from damaged mouse liver can be clonally expanded as organoids in Rspo1-based culture medium over several months. Such clonal organoids can be induced to differentiate in vitro and to generate functional hepatocytes upon transplantation into Fah(-/-) mice. These findings indicate that previous observations concerning Lgr5(+) stem cells in actively self-renewing tissues can also be extended to damage-induced stem cells in a tissue with a low rate of spontaneous proliferation.
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“Liver organoids derived from leucine-rich repeat–containing G protein–coupled receptor 5 (LGR5)–positive liver progenitor cells (LPC) can be stably maintained for months and can differentiate into functionally competent hepatocytes and cholangiocytes [14–16].”
From High-throughput human microfluidic organoid-on-a-chip platform for modeling liver diseases and screening nanotherapeutics. (2026) · cites this paper as [14]
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“The liver organoid has selfrenewal capability, a key proliferative property for its long-term culture and clonal expansion from single cells with sustained stemness (Huch et al., 2013;Hu et al., 2018).”
From Fast formation and maturation enhancement of human liver organoids using a liver-organoid-on-a-chip (2024) · cites this paper as Huch et al., 2013
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“These studies demonstrated that HPCs arise within the liver, engraft the parenchyma14 and may have some differentiation capacity but did not regenerate significant quantities of liver parenchyma11-13.”
From Hepatic progenitor cells of biliary origin with liver repopulation capacity (2015) · cites this paper as [12]
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Metadata
- Authors
- Huch, M., Dorrell, C., Boj, S. F., van Es, J. H., Li, V. S. W., van de Wetering, M., Sato, T., Hamer, K., Sasaki, N., Finegold, M. J., Haft, A., Vries, R. G., Grompe, M., & Clevers, H.
- Affiliations
- Utrecht University (NL)
- Oregon Health & Science University (US)
- Keio University (JP)
- Baylor College of Medicine (US)
- Corresponding author
- Clevers, Hans
- ORCID
- Venue
- Nature
- Published
- 2013-01-27
- Volume
- 494
- Issue
- 7436
- Pages
- 247-250
- ISSN
- 0028-0836 (ISSN-L)
- 1476-4687
- PMC
- PMC3634804
- OpenAlex
- W2008285918
- Cited by
- 1,555
- References
- 33
- Citations by year
- 2026: 66 · 2025: 117 · 2024: 148 · 2023: 118 · 2022: 122 · 2021: 143 · 2020: 146 · 2019: 120 · 2018: 125 · 2017: 120 · 2016: 109 · 2015: 80 · 2014: 95 · 2013: 43
- FWCI
- 67.87
- Type
- journal article
- Language
- en
- Open access
- green
- www.ncbi.nlm.nih.gov
- Locations
- Nature (publishedVersion)
- PubMed (publishedVersion)
- Europe PMC (PubMed Central) (acceptedVersion)
- PubMed Central (acceptedVersion)
- Data Archiving and Networked Services (DANS) (submittedVersion)
- RePEc: Research Papers in Economics (submittedVersion)
- KNAW Research Portal (The Royal Netherlands Academy of Arts and Sciences) (submittedVersion)
- Funders
- Wellcome Trust (104151)
- Japan Society for the Promotion of Science
- National Institute of Diabetes and Digestive and Kidney Diseases (R01DK051592, P30DK056338)
- Topics
- Health Sciences › Medicine › Hepatology › Liver physiology and pathology
- Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Wnt/β-catenin signaling in development and cancer
- Health Sciences › Medicine › Surgery › Tissue Engineering and Regenerative Medicine
- Keywords
- Wnt signaling, liver regeneration, liver organoids, hepatocyte differentiation, bile duct regeneration, tyrosinemia type I, RSPO1, clonal expansion, stem cell transplantation, liver injury, lineage tracing
- MeSH terms
- Alleles, Animals, Bile Ducts, Clone Cells, Culture Media, Disease Models, Animal, Female, Hydrolases, Liver, Liver Diseases, Male, Organoids, Regeneration, Stem Cells, Cell Lineage, Thrombospondins, Tyrosinemias, Hepatocytes, Multipotent Stem Cells, Receptors, G-Protein-Coupled, Mice, Gene Knock-In Techniques, Wnt Signaling Pathway
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