Explant culture of adult zebrafish hearts for epicardial regeneration studies.

Published

Journal Article

Here we describe how to culture adult zebrafish hearts as explants and study the regeneration of epicardial tissue ex vivo, as a means to identify therapeutic targets for heart disease. Uninjured or injured adult hearts are excised, washed and cultured in an incubator with gentle agitation. Heart explants can be prepared within 2 h, and they can be maintained in culture for 30 d or longer. If explants are prepared from appropriate transgenic lines, dynamic behaviors of epicardial cells can be monitored by live imaging using stereofluorescence microscopy. We also describe ex vivo procedures for genetic ablation of the epicardium, cell proliferation assays, tissue grafts and bead grafts. Basic cell culture and surgical skills are required to carry out this protocol. Unlike existing protocols for culturing isolated zebrafish epicardial cells on matrices, procedures described here maintain epicardial cells on an intact cardiac surface, thereby better supporting in vivo cell behaviors. Our protocols complement and extend in vivo studies of heart regeneration.

Full Text

Duke Authors

Cited Authors

  • Cao, J; Poss, KD

Published Date

  • May 2016

Published In

Volume / Issue

  • 11 / 5

Start / End Page

  • 872 - 881

PubMed ID

  • 27055096

Pubmed Central ID

  • 27055096

Electronic International Standard Serial Number (EISSN)

  • 1750-2799

International Standard Serial Number (ISSN)

  • 1754-2189

Digital Object Identifier (DOI)

  • 10.1038/nprot.2016.049

Language

  • eng