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  3. Preparation of chaperone-loaded neural stem cell-derived extracellular vesicles to reduce protein aggregation in Huntington's disease cellular models

Preparation of chaperone-loaded neural stem cell-derived extracellular vesicles to reduce protein aggregation in Huntington's disease cellular models

  • STAR Protoc. 2023 Mar 17;4(1):102134. doi: 10.1016/j.xpro.2023.102134.
Bhagyashree S Joshi 1,  Inge S Zuhorn 2
Affiliations

Affiliations

  • 1 Department of Biomedical Engineering, University of Groningen, University Medical Center Groningen, A. Deusinglaan 1, 9713 AV Groningen, the Netherlands. Electronic address: [email protected].
  • 2 Department of Biomedical Engineering, University of Groningen, University Medical Center Groningen, A. Deusinglaan 1, 9713 AV Groningen, the Netherlands. Electronic address: [email protected].
PMID: 36861837 DOI: 10.1016/j.xpro.2023.102134
Abstract

Here, we present a protocol using genetic engineering techniques to prepare small extracellular vesicles (sEVs) enriched in the chaperone protein DNAJB6. We describe steps to prepare cell lines overexpressing DNAJB6, followed by the isolation and characterization of sEVs from cell conditioned media. Further, we describe assays to examine effects of DNAJB6-loaded sEVs on protein aggregation in Huntington's disease cellular models. The protocol can be readily repurposed to study protein aggregation in Other neurodegenerative disorders or extended to Other therapeutic proteins. For complete details on the use and execution of this protocol, please refer to Joshi et al. (2021).1.

Keywords

Cell Biology; Cell Separation/Fractionation; Gene Expression; Health Sciences; Microscopy; Molecular Biology; Molecular/Chemical Probes; Neuroscience; Protein Biochemistry; Stem Cells.

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