Ruo, Matilde (2025) Dissociating and Accelerating the Maturation of Human Cortical Organoids. Research Project 1 (minor thesis), Behavioural and Cognitive Neurosciences.
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Abstract
Human brain organoids are a 3D in vitro model generated from human iPSCs, currently used to investigate brain-related diseases. Despite their potential, challenges such as long and complex culture protocols and slow maturation continue to limit their applicability in research. Therefore, we aim to improve organoid culturing, consistency, and yield by incorporating a mid-point cryopreservation step into the culturing protocol, and to enhance their applicability to aging research by accelerating organoid maturation. For the latter, we investigated the effects of two independent treatments targeting maturation-inducing epigenetic regulators on the maturation of cortical organoids, namely EZH2 inhibitor (EZH2i) and the GENtoniK cocktail of compounds. A dissociation protocol on day 20 and day 25 of the cortical organoid differentiation protocol was successfully implemented with both a direct reassociation and a cryopreservation step. Reassociated organoids were morphologically comparable to control organoids, as identified by H&E and immunofluorescence staining. As for the acceleration of organoid maturation, both EZH2i and GENtoniK treatments induced minimal changes in expression levels of neuronal marker genes, while astrocytic marker GFAP was significantly increased by both treatments. Hence, epigenetic regulators EZH2i and GENtoniK might influence astrocytic maturation in an organoid setting, although no convincing acceleration of neuronal maturation was observed.
| Item Type: | Thesis (Research Project 1 (minor thesis)) |
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| Supervisor name: | Kooistra, S.M. and Eggen, B.J.L. |
| Degree programme: | Behavioural and Cognitive Neurosciences |
| Thesis type: | Research Project 1 (minor thesis) |
| Language: | English |
| Date Deposited: | 15 Jul 2025 09:30 |
| Last Modified: | 15 Jul 2025 09:30 |
| URI: | https://fse.studenttheses.ub.rug.nl/id/eprint/35979 |
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