title: Reconstitution of a Patterned Neural Tube from Single Mouse Embryonic Stem Cells.
abstract: The recapitulation of tissue development and patterning in three-dimensional (3D) culture is an important dimension of stem cell research. Here, we describe a 3D culture protocol in which single mouse ES cells embedded in Matrigel under neural induction conditions clonally form a lumen containing, oval-shaped epithelial structure within 3 days. By Day 7 an apicobasally polarized neuroepithelium with uniformly dorsal cell identity forms. Treatment with retinoic acid at Day 2 results in posteriorization and self-organization of dorsal-ventral neural tube patterning. Neural tube organoid growth is also supported by pure laminin gels as well as poly(ethylene glycol) (PEG)-based artificial extracellular matrix hydrogels, which can be fine-tuned for key microenvironment characteristics. The rapid generation of a simple, patterned tissue in well-defined culture conditions makes the neural tube organoid a tractable model for studying neural stem cell self-organization.
authors.affiliation: DFG Research Center for Regenerative Therapies Dresden, Technische Universität Dresden, 01307, Dresden, Germany.
authors.forename: Keisuke
authors.initials: K
authors.lastname: Ishihara
authors.affiliation: Laboratory of Stem Cell Bioengineering, Institute of Bioengineering, School of Life Sciences and School of Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.|Department of Mechanical Engineering, KU Leuven, Belgium.
authors.forename: Adrian
authors.initials: A
authors.lastname: Ranga
authors.affiliation: Laboratory of Stem Cell Bioengineering, Institute of Bioengineering, School of Life Sciences and School of Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL), Lausanne, Switzerland.|Institute of Chemical Sciences and Engineering, School of Basic Sciences, EPFL, Lausanne, Switzerland.
authors.forename: Matthias P
authors.initials: MP
authors.lastname: Lutolf
authors.affiliation: Research Institute of Molecular Pathology, Campus Vienna Biocenter 1, 1030 Vienna, Austria. elly.tanaka@imp.ac.at.
authors.forename: Elly M
authors.initials: EM
authors.lastname: Tanaka
authors.affiliation: DFG Research Center for Regenerative Therapies Dresden, Technische Universität Dresden, 01307, Dresden, Germany. andrea.meinhardt@tu-dresden.de.
authors.forename: Andrea
authors.initials: A
authors.lastname: Meinhardt
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques / Q000379:methods; D002478:Cells, Cultured; D003094:Collagen / Q000008:administration & dosage; D004338:Drug Combinations; D047108:Embryonic Development / Q000187:drug effects / Q000502:physiology; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000187:drug effects; D005109:Extracellular Matrix / Q000502:physiology; D020100:Hydrogels / Q000008:administration & dosage; D007797:Laminin / Q000008:administration & dosage; D051379:Mice; D054259:Neural Tube / Q000166:cytology* / Q000187:drug effects; D038081:Organogenesis / Q000187:drug effects / Q000502:physiology*; D009940:Organoids / Q000166:cytology; D011092:Polyethylene Glycols / Q000008:administration & dosage; D011509:Proteoglycans / Q000008:administration & dosage; D014212:Tretinoin / Q000494:pharmacology
chemical_list: D004338:Drug Combinations; D020100:Hydrogels; D007797:Laminin; D011509:Proteoglycans; C059485:matrigel; D011092:Polyethylene Glycols; D014212:Tretinoin; D003094:Collagen
keywords: Artificial extracellular matrix; Cyst; Lumen; Mouse embryonic stem cells; Neural tube; Neuroepithelium; Organoid; PEG hydrogel; Patterning