title: Intestinal epithelial organoids fuse to form self-organizing tubes in floating collagen gels.
abstract: Multiple recent examples highlight how stem cells can self-organizein vitroto establish organoids that closely resemble theirin vivocounterparts. Single Lgr5+mouse intestinal stem cells can be cultured under defined conditions forming ever-expanding epithelial organoids that retain cell polarization, cell type diversity and anatomical organization of thein vivoepithelium. Although exhibiting a remarkable level of self-organization, the so called 'mini-guts' have a closed cystic structure of microscopic size. Here, we describe a simple protocol to generate macroscopic intestinal tubes from small cystic organoids. Embedding proliferating organoids within a contracting floating collagen gel allows them to align and fuse to generate macroscopic hollow structures ('tubes') that are lined with a simple epithelium containing all major cell types (including functional stem cells) of the small intestine. Cells lining the central contiguous lumen closely resemble the epithelial cells on luminal villiin vivo, whereas buds that protrude from the main tube into the surrounding matrix closely resemble crypts. Thus, the remarkable self-organizing properties of Lgr5+stem cells extend beyond the level of the microscopic cystic organoid to the next, macroscopic, level of tube formation.
authors.affiliation: Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW), and University Medical Center Utrecht, Uppsalalaan 8, Utrecht 3584 CT, The Netherlands.
authors.forename: Norman
authors.identifier: 0000-0002-5467-7151
authors.initials: N
authors.lastname: Sachs
authors.affiliation: Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW), and University Medical Center Utrecht, Uppsalalaan 8, Utrecht 3584 CT, The Netherlands.
authors.forename: Yoshiyuki
authors.initials: Y
authors.lastname: Tsukamoto
authors.affiliation: The Netherlands Cancer Institute, Antoni van Leeuwenhoek Hospital, Plesmanlaan 121, Amsterdam 1066 CX, The Netherlands.
authors.forename: Pekka
authors.initials: P
authors.lastname: Kujala
authors.affiliation: The Maastricht Multimodal Molecular Imaging Institute, Universiteitssingel 50, Maastricht 6229 ER, The Netherlands.
authors.forename: Peter J
authors.identifier: 0000-0002-2964-5684
authors.initials: PJ
authors.lastname: Peters
authors.affiliation: Hubrecht Institute, Royal Netherlands Academy of Arts and Sciences (KNAW), and University Medical Center Utrecht, Uppsalalaan 8, Utrecht 3584 CT, The Netherlands h.clevers@hubrecht.eu.
authors.forename: Hans
authors.identifier: 0000-0002-3077-5582
authors.initials: H
authors.lastname: Clevers
mesh_terms: D000818:Animals; D002454:Cell Differentiation / Q000187:drug effects; D002459:Cell Fusion; D003094:Collagen / Q000494:pharmacology*; D005782:Gels / Q000494:pharmacology*; D007413:Intestinal Mucosa / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D051379:Mice; D009940:Organoids / Q000166:cytology* / Q000187:drug effects / Q000648:ultrastructure; D051381:Rats; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects; D046509:Tissue Culture Techniques / Q000379:methods*
chemical_list: D005782:Gels; D003094:Collagen
keywords: Collagen gel; Matrigel; Mini-gut; Mouse; Organoid; Tube