PhD Scientific Days 2024

Budapest, 9-10 July 2024

Poster Session D - Neurosciences 1.

Development of a Human Induced Pluripotent Stem Cell-Derived Cerebellar Organoid Model


Alexandra Sándor1, Kornélia Szebényi1, Katalin Monostory1
1: Institute of Molecular Life Science, HUN-REN Research Centre for Natural Sciences

Text of the abstract

The cerebellum is important for processing motor and sensory information. Due to its continoued development during the post-natal period, it is vulnerable to a number of pathological processes, for example to different type of ataxias. Since the development of the human cerebellum is completely different from that of the mouse cerebellum, it is essential to use human model systems to study both developmental and neurodegenerative processes.
We have previously developed a human induced pluripotent stem cell (hiPSC)-derived cerebellar organoid system, containingthe disease relevant cell types in a tissue-like organization. However, Calbindin-positive Purkinje neurons seen on day 50 of the organoid differentiation do not show mature Purkinje morphology.
Increasing the duration of cerebellar organoid culture times can lead to the generation of Purkinje neurons with morphology and functions similar to postnatal cells. From the point of view of disease modeling, this provides a more relevant test system for the development of therapeutic interventions than our currently available cerebellar organoid model.
Therefore we increased the culture times of the cerebellar organoids from 50 days to 120 days to develope a reproducible differentiation protocol for the generation of a hiPSC-derived organoid model of the cerebellum containing mature Purkinje neurons. From day 75 to day 120, the organoids were maintained in two different conditions, where one of the mediums has been supplemented with glucose. The cell types found in the organoids as well as the morphology of Purkinje neurons on days 75 and 120 of the differentiation were characterized by immunocytochemical methods.
Overall, it can be concluded that we have created an organoid model of the human cerebellum, which contains mature Purkinje neurons, so in the future, we can use this improved protocol to study neurodegenerative diseases.
I am deeply grateful to the ÚNKP (23-3-I.) for their generous support, which has been instrumental in both the progress of my research and the creation of the poster presentation. Their funding has significantly contributed to the success of my work, and I appreciate the opportunities it has provided.