Explant culture of adult zebrafish hearts for epicardial regeneration studies.
Journal Article (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
- PMC4857716
Electronic International Standard Serial Number (EISSN)
- 1750-2799
Digital Object Identifier (DOI)
- 10.1038/nprot.2016.049
Language
- eng
Conference Location
- England