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.
Associated documents
Links
Luminescent Human iPSC-Derived Neurospheroids Enable Modeling of Neurotoxicity …

Contact person

Elise Van Breedam

Organisations

University of Antwerp (UAntwerpen)
Vaccine and Infectious Disease Institute (Vaxinfectio)
Belgium