Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium

Toshiro Sato, Daniel E. Stange, Marc Ferrante, Robert G J Vries, Johan H. Van Es, Stieneke Van Den Brink, Winan J. Van Houdt, Apollo Pronk, Joost Van Gorp, Peter D. Siersema, Hans Clevers

Research output: Contribution to journalArticle

934 Citations (Scopus)

Abstract

Background & Aims: We previously established long-term culture conditions under which single crypts or stem cells derived from mouse small intestine expand over long periods. The expanding crypts undergo multiple crypt fission events, simultaneously generating villus-like epithelial domains that contain all differentiated types of cells. We have adapted the culture conditions to grow similar epithelial organoids from mouse colon and human small intestine and colon. Methods: Based on the mouse small intestinal culture system, we optimized the mouse and human colon culture systems. Results: Addition of Wnt3A to the combination of growth factors applied to mouse colon crypts allowed them to expand indefinitely. Addition of nicotinamide, along with a small molecule inhibitor of Alk and an inhibitor of p38, were required for long-term culture of human small intestine and colon tissues. The culture system also allowed growth of mouse Apc-deficient adenomas, human colorectal cancer cells, and human metaplastic epithelia from regions of Barrett's esophagus. Conclusions: We developed a technology that can be used to study infected, inflammatory, or neoplastic tissues from the human gastrointestinal tract. These tools might have applications in regenerative biology through ex vivo expansion of the intestinal epithelia. Studies of these cultures indicate that there is no inherent restriction in the replicative potential of adult stem cells (or a Hayflick limit) ex vivo.

Original languageEnglish
Pages (from-to)1762-1772
Number of pages11
JournalGastroenterology
Volume141
Issue number5
DOIs
Publication statusPublished - 2011 Nov
Externally publishedYes

Fingerprint

Organoids
Barrett Esophagus
Adenoma
Colon
Adenocarcinoma
Small Intestine
Adult Stem Cells
Niacinamide
Intestinal Mucosa
Gastrointestinal Tract
Colorectal Neoplasms
Intercellular Signaling Peptides and Proteins
Stem Cells
Epithelium
Technology
Growth

Keywords

  • Dysplasia
  • Ex Vivo Culture Technology
  • Hayflick Model
  • Neoplasia
  • Signaling

ASJC Scopus subject areas

  • Gastroenterology

Cite this

Sato, T., Stange, D. E., Ferrante, M., Vries, R. G. J., Van Es, J. H., Van Den Brink, S., ... Clevers, H. (2011). Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. Gastroenterology, 141(5), 1762-1772. https://doi.org/10.1053/j.gastro.2011.07.050

Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. / Sato, Toshiro; Stange, Daniel E.; Ferrante, Marc; Vries, Robert G J; Van Es, Johan H.; Van Den Brink, Stieneke; Van Houdt, Winan J.; Pronk, Apollo; Van Gorp, Joost; Siersema, Peter D.; Clevers, Hans.

In: Gastroenterology, Vol. 141, No. 5, 11.2011, p. 1762-1772.

Research output: Contribution to journalArticle

Sato, T, Stange, DE, Ferrante, M, Vries, RGJ, Van Es, JH, Van Den Brink, S, Van Houdt, WJ, Pronk, A, Van Gorp, J, Siersema, PD & Clevers, H 2011, 'Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium', Gastroenterology, vol. 141, no. 5, pp. 1762-1772. https://doi.org/10.1053/j.gastro.2011.07.050
Sato, Toshiro ; Stange, Daniel E. ; Ferrante, Marc ; Vries, Robert G J ; Van Es, Johan H. ; Van Den Brink, Stieneke ; Van Houdt, Winan J. ; Pronk, Apollo ; Van Gorp, Joost ; Siersema, Peter D. ; Clevers, Hans. / Long-term expansion of epithelial organoids from human colon, adenoma, adenocarcinoma, and Barrett's epithelium. In: Gastroenterology. 2011 ; Vol. 141, No. 5. pp. 1762-1772.
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