Development of luminescent human iPSC-derived neurospheroids
Scope of the method
The Method relates to
- Human health
The Method is situated in
- Basic Research
Type of method
- In vitro - Ex vivo
This method makes use of
- Human derived cells / tissues / organs
Specify the type of cells/tissues/organs
human induced pluripotent stem cell-derived neurospheroids
Description
Method keywords
- neurospheroid
- Bioluminescence
- IPSC
- organoid
- neurotoxicity
Scientific area keywords
- 3D organoid models
- Induced pluripotent stem cells
- ischemic stroke
Method description
This method relates to the development of highly reproducible human iPSC-derived neurospheroids equipped with intrinsic bioluminescence for an easy and longitudinal follow-up of the viability and growth of these neurospheroids over time. The luminescent neurospheroids have been applied in ischemic stroke research, where this model enabled modeling of neurotoxicity after oxygen-glucose deprivation. The easy neural survival read-out may also enable the evaluation of potential neuroprotective agents (in high-throughput).
Lab equipment
- - Laminar flow cabinet;
- - Shaker;
- - Microplate reader (Luminometer).
Method status
- Published in peer reviewed journal
Pros, cons & Future potential
Advantages
- - Three-dimensional model;
- - human-based model;
- - longitudinal measurements of neurospheroid viability (i.e. does not require a single endpoint and/or disruption of neurospheroids);
- - highly reproducible;
- - amenable to high-throughput drug screening.
Challenges
- - Maturity of neurospheroids and lack of glial cell types;
- - Hypoxic/necrotic core development;
- - Potential transgene silencing associated with lentiviral vector transduction.
Modifications
- - Optimization of culture conditions of neurospheroids (i.e. increasing culture time, other media types, use of bioreactors, etc.);
- - Addition of microglia-progenitors to neurospheroids;
- - Modification of genetic engineering strategy (e.g. CRISPR/Cas9).
Future & Other applications
- - Neurotoxicity, neurotrauma and neurodegenerative disease modeling;
- - Evaluation of candidate neuroprotective therapies (in high-throughput).
References, associated documents and other information
References
- Van Breedam E, Nijak A, Buyle-Huybrecht T, Di Stefano J, Boeren M, Govaerts J, et al. Luminescent Human iPSC-Derived Neurospheroids Enable Modeling of Neurotoxicity After Oxygen-glucose Deprivation. Neurotherapeutics. 2022.
Links
Luminescent Human iPSC-Derived Neurospheroids Enable Modeling of Neurotoxicity …
Contact person
Elise Van BreedamOrganisations
University of Antwerp (UAntwerpen)Vaccine and Infectious Disease Institute (Vaxinfectio)
Belgium
Flemish Region