Engineering CNS Tissue Morphogenesis Ex Vivo

The human central nervous system (CNS) is comprised of diverse interconnected tissues, each containing unique cytoarchitectures and cell phenotypes critical to their physiology and associated disease pathologies. Engineering organotypic models of these tissues from human pluripotent stem cells (hSPCs) poses unique challenges because the morphogenetic events that pattern CNS development in vivo are dynamic, combinatorial, and occur at micro- to-millimeter scales. Organoid models to date rely on uncontrolled, spontaneous morphogenesis of EBs or neurospheres that provide limited to no control over the resulting tissue cytoarchitecture. Here, we present novel culture platforms that enable the requisite spatial and temporal control of microscale morphogen gradients and tissue morphology necessary to orchestrate morphogenesis of 2D transverse slice and 3D tube models of the developing spinal cord. These platforms integrate our latest advancements in chemically defined neural
differentiation protocols1,2 with micropatterned culture substrates3,4 and sacrificial template-molded hydrogels.
Furthermore, we will describe our advances in implementing such platforms to create a high throughput screening platform for developmental neurotoxicity studies and neurodegenerative disease models.
References:
1. Lippmann, E. S. et al. Stem Cells 32, 1032–1042 (2014).
2. Lippmann, E. S. et al. Stem Cell Reports 4, 632–644 (2015).
3. Knight, G. T. et al. Chem. Commun. (Camb.) 51, 5238–5241 (2015).
4. McNulty, J. D. et al. Lab Chip 14, 1923–1930 (2014).