Development of Myelinating Human Brain Organoids for Modelling Multiple Sclerosis: A Platform For Drug Screening Studies and Mechanistic Investigations

Year 2025, Volume: 4 Issue: 2, 64 - 76, 13.12.2025

Büşra Acar , Alaattin Şen

https://doi.org/10.5281/zenodo.17786373

Abstract

Multiple sclerosis (MS) is a neurodegenerative disease that is characterized mainly by demyelination and inflammation. The disadvantages of the current two-dimensional cell culture models and experimental animals imply the need for the development of new disease models. Brain organoids are emerging and powerful tools for recapitulating the central nervous system. Although the models have been generated for many years, time-consuming processes and less cellular diversity are important challenges in defined protocols. In this study, we aimed to generate an optimized brain organoid protocol and model organoids for MS. In these processes, human embryonic stem cells were directed to neurons via the extracellular matrix (ECM) and growth factors to generate brain organoids with diverse neural cells. The presence of myelinating cells, astrocytes, microglia and excitatory neurons in the organoids was verified by immunostaining. MS was then used to induce inflammation and demyelination via lipopolysaccharide (LPS). The model was also confirmed by immunostaining, which revealed an ~50% increase in GFAP and an ~60% decrease in CNPase-positive cells. Finally, the use of organoids in the drug screening field was tested via fingolimod treatment of LPS-induced organoids. A comparison between fingolimod-treated and untreated organoids revealed that fingolimod decreased GFAP by more than 80% and increased the percentage of CNPase-positive cells by 90%. Additionally, the relative expression levels of inflammation-related transcripts (FOXP3 and GFAP) after fingolimod treatment were significantly decreased. In conclusion, a human brain organoid model for MS studies was successfully generated for use in drug screening and mechanistic studies.

Keywords

brain organoids , myelination , inflammation , multiple sclerosis

Ethical Statement

We declare that the manuscript that above information does not require any “Ethics Committee Permission”; it is an original work; compliance with scientific ethical principles and rules is observed in all stages of preparation, data collection, analysis and presentation of information; all data and information not obtained within the scope of the study are cited and that these sources are included in the references; accepts that no changes have been made to the data used, that all ethical terms and conditions specified in RESPECT's EU Code of Ethics for Socio-Economic Research, Committee on Publication Ethics (COPE) and Higher Education Institutions Scientific Research and Publication Ethics Directive are accepted all ethical duties and responsibilities are complied.

Supporting Institution

TUBİTAK

Project Number

TUBITAK 119Z389

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