In vitro expansion of single Lgr5+ liver stem cells induced by Wnt-driven regeneration

By Meritxell Huch, C. Dorrell, S. Boj, Johan H. van Es, M. van de Wetering, Vivian S. W. Li, K. Hamer, Nobuo Sasaki, M. Finegold, Annelise Haft and 2 more. Nature, 2013

1,454 citations recorded by the source corpus.

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.

Subfields: Hepatology, Molecular Biology, Surgery

View Paper: https://www.semanticscholar.org/paper/fbbd3f3cec524f7716ac714376e89f96e93342fd

How researchers see this paper

The researchers whose trusted networks rank it highest, strongest advocate first. Every researcher gets a different order.

In Hepatology

1. Hans Clevers
2. Charles M. Rice

In Molecular Biology

1. Hans Clevers
2. R. Nusse
3. W. de Lau
4. W. Peng
5. P. Gros
6. Juergen A. Knoblich
7. Toshiro Sato
8. M. van de Wetering

In Surgery

1. Hans Clevers
2. R. Nusse
3. W. de Lau
4. W. Peng
5. P. Gros

Citations in the corpus

• Modeling Development and Disease with Organoids. (2016, background): “Clonal pancreas organoids differentiated along ductal and endocrine lineages when grafted Cell 165, June 16, 2016 1591 in vivo in a developing pancreas, indicative of bipotentiality (Huch et al., 2013a).”
• The R-spondin/Lgr5/Rnf43 module: regulator of Wnt signal strength (2014, positive): “…lineage tracing strategy, Lgr5 was subsequently found to mark stem cells in many other organs and tissues, including the stomach (Barker et al. 2010), pancreas (Huch et al. 2013a), liver (Huch et al. 2013b), kidney (Barker et al. 2012), and mammary gland (de Visser et al. 2012; Plaks et al. 2013).”
• Microfluidic 3D cell culture: from tools to tissue models. (2015, background): “Stem cells expressing the LGR5 receptor have been identified which have been shown to maintain stemness in vitro and differentiate into fully functional intestine, stomach and liver [56–58].”
• Lessons from the cancer genome. (2013, neutral): “However, new methods (such as Rho kinase inhibitor-treated feeder layers and ‘‘organoid’’ culture systems) appear poised to greatly expand the repertoire of available cancer models (Huch et al., 2013; Liu et al., 2012).”
• Unlimited in vitro expansion of adult bi-potent pancreas progenitors through the Lgr5/R-spondin axis (2013, positive): “We have recently described a 3D culture system that allows long-term expansion of adult small intestine, stomach and liver cells without the need of a mesenchymal niche, while preserving the characteristics of the original adult epithelium (Sato et al , 2009; Barker et al , 2010; Huch et al , 2013).”
• An integral program for tissue renewal and regeneration: Wnt signaling and stem cell control (2014, background): “Comparable protocols have been established for Lgr5 cells derived from the stomach (85), liver (86), and pancreas (87).”
• Long-Term Expansion of Functional Mouse and Human Hepatocytes as 3D Organoids. (2018, positive): “Indeed, oval cells or atypical ductal cells expressing Epcam or Sox9 have long been regarded as bi-potential facultative stem/ progenitor cells (Dolle´ et al., 2015; Furuyama et al., 2011; Huch and Clevers, 2011; Huch et al., 2013).”
• Identification of multipotent luminal progenitor cells in human prostate organoid cultures (2014, background): “Organoids remain genetically and phenotypically stable in culture, exemplified by pathology-free transplantation ofmultiplemicewith the organoid offspring of single Lgr5+ cells from colon (Yui et al., 2012) or liver (Huch et al., 2013).”
• Effects of diurnal variation of gut microbes and high fat feeding on host circadian clock function and metabolism (2015, methodology): “Hepatic organoids (Huch et al., 2013) (hepanoids) were established in vitro and their CC cell synchronization was initiated via serum shock (Balsalobre et al, 1998).”
• Organoid Models of Human and Mouse Ductal Pancreatic Cancer (2014, background): “…organoids by modifying approaches we previously pioneered to culture intestinal (Sato et al., 2009), gastric (Barker et al., 2010), colon carcinoma (Sato et al., 2011), hepatic (Huch et al., 2013b), pancreatic (Huch et al., 2013a), and prostatic organoids (Gao et al., 2014; Karthaus et al., 2014).”