Imaging Autophagy in hiPSC-Derived Midbrain Dopaminergic Neuronal Cultures for Parkinson’s Disease Research

Petros Stathakos, Natalia Jimenez Moreno, Lucy Crompton, Paul Nistor, Maeve Caldwell, Jon Lane

Research output: Chapter in Book/Report/Conference proceedingChapter in a book

1 Citation (Scopus)
95 Downloads (Pure)

Abstract

To appreciate the positive or negative impact of autophagy during the initiation and progression of human diseases, the isolation or de novo generation of appropriate cell types is required to support focused in vitro assays. In human neurodegenerative diseases such as Parkinson’s disease (PD), specific subsets of acutely sensitive neurons become susceptible to stress-associated operational decline and eventual cell death, emphasizing the need for functional studies in those vulnerable groups of neurons. In PD, a class of dopaminergic neurons in the ventral midbrain (mDANs) is affected. To study these, human-induced pluripotent stem cells (hiPSCs) have emerged as a valuable tool, as they enable the establishment and study of mDAN biology in vitro. In this chapter, we describe a stepwise protocol for the generation of mDANs from hiPSCs using a monolayer culture system. We then outline how imaging-based autophagy assessment methodologies can be applied to these neurons, thereby providing a detailed account of the application of imaging-based autophagy assays to human iPSC-derived mDANs
Original languageEnglish
Title of host publicationAutophagy
PublisherHumana Press
Pages257-280
Number of pages23
DOIs
Publication statusPublished - 5 Jan 2019

Publication series

NameMethods in Molecular Biology
PublisherSpringer
ISSN (Electronic)1064-3745

Keywords

  • Autophagy
  • Parkinson’s disease
  • Immunofluorescence
  • Cell culture
  • Dopaminergic neurons
  • Stem cells
  • hiPSC

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