pp 1-17

Part of the Methods in Molecular Biology book series | Cite as

Disaggregation and Reaggregation of Zebrafish Retinal Cells for the Analysis of Neuronal Layering

  • Megan K. Eldred
  • Leila Muresan
  • William A. Harris
Protocol

Abstract

The reaggregation of dissociated cells to form organotypic structures provides an in vitro system for the analysis of the cellular interactions and molecular mechanisms involved in the formation of tissue architecture. The retina, an outgrowth of the forebrain, is a precisely layered neural tissue, yet the mechanisms underlying layer formation are largely unexplored. Here we describe the protocol to dissociate, re-aggregate, and culture zebrafish retinal cells from a transgenic, Spectrum of Fates, line where all main cell types are labelled with a combination of fluorescent proteins driven by fate-specific promoters. These cells re-aggregate and self-organize in just 48 h in minimal culture conditions. We also describe how the patterning in these aggregates can be analyzed using isocontour profiling to compare whether different conditions affect their self-organization.

Keywords:

3D Petri dish Aggregation Isocontour profiling Müller Glia Organoid Retina Self-organization SoFa Zebrafish 

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Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Megan K. Eldred
    • 1
  • Leila Muresan
    • 1
  • William A. Harris
    • 1
  1. 1.Department of Physiology, Development and NeuroscienceCambridge UniversityCambridgeUK

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