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

doi:10.1016/j.xpro.2024.102875

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

STAR Protoc. 2024 Mar 15;5(1):102875. doi: 10.1016/j.xpro.2024.102875. Epub 2024 Feb 22.

Authors

Aikaterini A Kalargyrou¹, Ayako Matsuyama², Emily P Lanning², Mahmoud Khazim², Siobhan Guilfoyle², Alexander J Smith², Robin R Ali², Rachael A Pearson³

Affiliations

¹ Centre for Gene Therapy and Regenerative Medicine, King's College London, Guy's Hospital, 8th Floor Tower Wing, London SE1 9RT, UK. Electronic address: akalargyrou@meei.harvard.edu.
² Centre for Gene Therapy and Regenerative Medicine, King's College London, Guy's Hospital, 8th Floor Tower Wing, London SE1 9RT, UK.
³ Centre for Gene Therapy and Regenerative Medicine, King's College London, Guy's Hospital, 8th Floor Tower Wing, London SE1 9RT, UK. Electronic address: rachael.pearson@kcl.ac.uk.

Abstract

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.

Keywords: Cell Biology; Molecular Biology; Neuroscience.

MeSH terms

Animals
Cell Culture Techniques
Dissection
Extracellular Vesicles*
Mice
Retina*