Towards engineering an artificial neural tube

In this thesis, I present a self-organizing neural tube organoid that is strikingly similar in morphology, cell-type composition, and patterning to the mouse embryonic neural tube. When exposed to a sequence of epiblast culture conditions and neural differentiation conditions in 3D Matrigel, single mouse embryonic stem cells develop into spontaneously elongating neuroepithelial tissues organized with key hallmarks of the neural tube. Single cell transcriptomics revealed the presence of regionalized cell types spanning the region from the midbrain to the spinal cord along the AP axis of the embryo. In long-term culture experiments for organoid maturation, I observed the emergence of neural crest cells and mature neurons. Furthermore, I developed a microfluidics-based system to promote a similar DV patterning to a neural tube. Also, maturation of neural tube organoid exhibited the emergence and migration of multipotent neural crest cells. Collectively, the accessibility and scalability of these organoids make them a unique model of the developing neural tube, allowing the study of key mechanisms involved in striking morphogenesis by self-organization, cell regionalization, neural crest development, and morphogen-triggered patterning in vivo.