Poster presented at ISSCR 2022 by Seungmi Ryu. The poster is entitled: Generating Cerebellar Organoids from Human Pluripotent Stem Cells.

Authors (underlined, presenting): Seungmi Ryu, Hyenjong Hong, Vukasin M. Jovanovic, Carlos A. Tristan, Anton Simeonov, Ilyas Singeç

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Abstract

The cerebellum plays an important role in motor coordination and is affected in various neurological conditions. Most of our current understanding of the human cerebellum is derived from postmortem tissues, animal models, and immortalized cell lines cultured in conventional 2D cultures. Over the past decade, human induced pluripotent stem cells (iPSCs) have become increasingly popular for generating various 3D organoid models. However, little is known about how to generate bona fide cerebellar tissue from iPSCs. Here, we developed a novel 60-day organoid differentiation strategy that recapitulates cerebellum-specific developmental processes. We demonstrate the generation of two distinct primordial regions that resemble the rhombic lip and cerebellar plate ventricular zone. Immunostainings and Western blot analysis confirmed that these primordial regions give rise to major neuronal cell
type of the cerebellum (e.g., Purkinje neuron-like cells, granule cells, interneurons) and Bergmann glia-like cells that are specialized to guide migratory granule cells to their final destination. Remarkably, self-organizing cerebellar organoids formed the appropriate layers of the cerebellum and neuronal migration followed an outside-in pattern, which contrasts with telencephalic brain regions and organoids that follow an inside-out migration pattern. To demonstrate utility for disease modelling, cerebellar organoids were generated using iPSCs derived from patients with Friedreich’s ataxia, an autosomal-recessive neurogenerative disorder caused by a defect in the frataxin gene. Ongoing molecular, cellular, and electrophysiological studies are aimed at elucidating disease phenotypes and testing therapeutic modalities including drug screening and gene editing with CRISPR-Cas9. In summary, our organoid model provides new opportunities to study the cerebellum in basic and translation research with potential future applications in regenerative medicine.