Novel bioengineering strategies for modeling cerebellar ataxias

Teresa Maria Pereira da Silva

Key information

Authors:

Teresa Maria Pereira da Silva (Teresa Maria Pereira da Silva)

Supervisors:

Maria do Carmo Salazar Velez Roque da Fonseca; Evguenia Pavlovna Bekman (Evguenia Pavlovna Bekman Gaspar); Tiago Paulo Gonçalves Fernandes (Tiago Paulo Gonçalves Fernandes)

Published in

10/26/2018

Abstract

The cerebellum plays a critical role in all vertebrates, and many neurological disorders such as ataxias are associated with cerebellar dysfunction. A major limitation in cerebellar research has been the lack of adequate disease models. Human induced pluripotent stem cells (iPSCs) have great potential for disease modeling and provide a valuable source for regenerative approaches. Therefore, the main objective of this thesis was to create new approaches for modeling cerebellar ataxias using iPSC technology. Firstly, a highly reproducible methodology for generating cerebellar neurons under chemically defined and feeder-free culture conditions was established. Besides that, the manipulation of different signaling pathways was performed in order to achieve a higher number of GABAergic progenitors and functional Purkinje cells, which are important cerebellar neurons involved in cerebellar dysfunction. After the establishment of a new protocol for cerebellar differentiation, scalable generation of human iPSC-derived organoids using the novel PBS 0.1 (100 mL) Vertical-Wheel single-use bioreactors was implemented. In this system, human iPSC-derived organoids were able to adopt cerebellar identity and mature into different cerebellar neurons and to survive for up to 3 months, without feeder layers.