Reconstitution of a Patterned Neural Tube from Single Mouse Embryonic Stem Cells

Part of the Methods in Molecular Biology book series (MIMB, volume 1597)


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.

Key words

Mouse embryonic stem cells Organoid Neural tube Neuroepithelium Cyst Lumen PEG hydrogel Artificial extracellular matrix Patterning 



This work was supported by a seed grant and core support from the DFG Research Center of Regenerative Therapies Dresden, the International Foundation for Paraplegia, and the Saw-2011-IPF-2 68 grant of the Leibniz Society, the BMBF (Systems Biology), EU framework 7 HEALTH research programme PluriMes (, an ERC grant (StG_311422), and funding from FZ111/EXC168. KI was supported by the ELBE fellowship from the Center for Systems Biology Dresden.


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Copyright information

© Springer Science+Business Media LLC 2017

Authors and Affiliations

  1. 1.DFG Research Center for Regenerative Therapies DresdenTechnische Universität DresdenDresdenGermany
  2. 2.Laboratory of Stem Cell BioengineeringInstitute of Bioengineering, School of Life Sciences and School of Engineering, Ecole Polytechnique Fédérale de Lausanne (EPFL)LausanneSwitzerland
  3. 3.Institute of Chemical Sciences and Engineering, School of Basic Sciences, EPFLLausanneSwitzerland
  4. 4.Research Institute of Molecular Pathology1030 ViennaAustria
  5. 5.Department of Mechanical EngineeringKU LeuvenBelgium

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