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.
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Acknowledgments
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.
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Ishihara, K., Ranga, A., Lutolf, M.P., Tanaka, E.M., Meinhardt, A. (2017). Reconstitution of a Patterned Neural Tube from Single Mouse Embryonic Stem Cells. In: Tsuji, T. (eds) Organ Regeneration. Methods in Molecular Biology, vol 1597. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6949-4_4
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DOI: https://doi.org/10.1007/978-1-4939-6949-4_4
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