STAR Protocols (Mar 2024)

A protocol for isolation and culturing of mouse primary postmitotic photoreceptors and isolation of extracellular vesicles

  • Aikaterini A. Kalargyrou,
  • Ayako Matsuyama,
  • Emily P. Lanning,
  • Mahmoud Khazim,
  • Siobhan Guilfoyle,
  • Alexander J. Smith,
  • Robin R. Ali,
  • Rachael A. Pearson

Journal volume & issue
Vol. 5, no. 1
p. 102875

Abstract

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Summary: Here, we present a protocol for isolating and culturing mouse photoreceptors in a minimal, chemically defined medium free from serum. We describe steps for retina dissection, enzymatic dissociation, photoreceptor enrichment, cell culture, extracellular vesicles (EVs) enrichment, and EV ultrastructural analysis. This protocol, which has been verified for cultured cells derived from multiple murine strains, allows for the study of several aspects of photoreceptor biology, including EV isolation and nanotube formation.For complete details on the use and execution of this protocol, please refer to Kalargyrou et al. (2021).1 : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.

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