Reconstitution of a Patterned Neural Tube from Single Mouse Embryonic Stem Cells
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 wordsMouse 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 (http://www.plurimes.eu/), an ERC grant (StG_311422), and funding from FZ111/EXC168. KI was supported by the ELBE fellowship from the Center for Systems Biology Dresden.
- 2.Kelava I, Lancaster MA (2016) Stem cell models of human brain development. Stem Cell 18:736–748Google Scholar