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

Meritxell Huch, Craig Dorrell, Sylvia F. Boj, Johan H. Van Es, Vivian S W Li, Marc Van De Wetering, Toshiro Sato, Karien Hamer, Nobuo Sasaki, Milton J. Finegold, Annelise Haft, Robert G. Vries, Markus Grompe, Hans Clevers

Research output: Contribution to journalArticle

560 Citations (Scopus)

Abstract

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.

Original languageEnglish (US)
Pages (from-to)247-250
Number of pages4
JournalNature
Volume494
Issue number7436
DOIs
StatePublished - Feb 14 2013

Fingerprint

Organoids
Regeneration
Stem Cells
Liver
Bile Ducts
Culture Media
Hepatocytes
Hair Follicle
G-Protein-Coupled Receptors
Leucine
Small Intestine
Cysts
Stomach
Colon
Transplantation
Alleles
Ligands
In Vitro Techniques
Genes

ASJC Scopus subject areas

  • General

Cite this

Huch, M., Dorrell, C., Boj, S. F., Van Es, J. H., Li, V. S. W., Van De Wetering, M., ... Clevers, H. (2013). In vitro expansion of single Lgr5 + liver stem cells induced by Wnt-driven regeneration. Nature, 494(7436), 247-250. https://doi.org/10.1038/nature11826

In vitro expansion of single Lgr5 + liver stem cells induced by Wnt-driven regeneration. / Huch, Meritxell; Dorrell, Craig; Boj, Sylvia F.; Van Es, Johan H.; Li, Vivian S W; Van De Wetering, Marc; Sato, Toshiro; Hamer, Karien; Sasaki, Nobuo; Finegold, Milton J.; Haft, Annelise; Vries, Robert G.; Grompe, Markus; Clevers, Hans.

In: Nature, Vol. 494, No. 7436, 14.02.2013, p. 247-250.

Research output: Contribution to journalArticle

Huch, M, Dorrell, C, Boj, SF, Van Es, JH, Li, VSW, Van De Wetering, M, Sato, T, Hamer, K, Sasaki, N, Finegold, MJ, Haft, A, Vries, RG, Grompe, M & Clevers, H 2013, 'In vitro expansion of single Lgr5 + liver stem cells induced by Wnt-driven regeneration', Nature, vol. 494, no. 7436, pp. 247-250. https://doi.org/10.1038/nature11826
Huch M, Dorrell C, Boj SF, Van Es JH, Li VSW, Van De Wetering M et al. In vitro expansion of single Lgr5 + liver stem cells induced by Wnt-driven regeneration. Nature. 2013 Feb 14;494(7436):247-250. https://doi.org/10.1038/nature11826
Huch, Meritxell ; Dorrell, Craig ; Boj, Sylvia F. ; Van Es, Johan H. ; Li, Vivian S W ; Van De Wetering, Marc ; Sato, Toshiro ; Hamer, Karien ; Sasaki, Nobuo ; Finegold, Milton J. ; Haft, Annelise ; Vries, Robert G. ; Grompe, Markus ; Clevers, Hans. / In vitro expansion of single Lgr5 + liver stem cells induced by Wnt-driven regeneration. In: Nature. 2013 ; Vol. 494, No. 7436. pp. 247-250.
@article{141c05fbb03c427baf090c806b64a3fd,
title = "In vitro expansion of single Lgr5 + liver stem cells induced by Wnt-driven regeneration",
abstract = "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.",
author = "Meritxell Huch and Craig Dorrell and Boj, {Sylvia F.} and {Van Es}, {Johan H.} and Li, {Vivian S W} and {Van De Wetering}, Marc and Toshiro Sato and Karien Hamer and Nobuo Sasaki and Finegold, {Milton J.} and Annelise Haft and Vries, {Robert G.} and Markus Grompe and Hans Clevers",
year = "2013",
month = "2",
day = "14",
doi = "10.1038/nature11826",
language = "English (US)",
volume = "494",
pages = "247--250",
journal = "Nature",
issn = "0028-0836",
publisher = "Nature Publishing Group",
number = "7436",

}

TY - JOUR

T1 - In vitro expansion of single Lgr5 + liver stem cells induced by Wnt-driven regeneration

AU - Huch, Meritxell

AU - Dorrell, Craig

AU - Boj, Sylvia F.

AU - Van Es, Johan H.

AU - Li, Vivian S W

AU - Van De Wetering, Marc

AU - Sato, Toshiro

AU - Hamer, Karien

AU - Sasaki, Nobuo

AU - Finegold, Milton J.

AU - Haft, Annelise

AU - Vries, Robert G.

AU - Grompe, Markus

AU - Clevers, Hans

PY - 2013/2/14

Y1 - 2013/2/14

N2 - 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.

AB - 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.

UR - http://www.scopus.com/inward/record.url?scp=84873712443&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=84873712443&partnerID=8YFLogxK

U2 - 10.1038/nature11826

DO - 10.1038/nature11826

M3 - Article

VL - 494

SP - 247

EP - 250

JO - Nature

JF - Nature

SN - 0028-0836

IS - 7436

ER -