PUBLICATION
Explant culture of adult zebrafish hearts for epicardial regeneration studies
- Authors
- Cao, J., Poss, K.D.
- ID
- ZDB-PUB-160408-3
- Date
- 2016
- Source
- Nature Protocols 11: 872-881 (Journal)
- Registered Authors
- Cao, Jingli, Poss, Kenneth D.
- Keywords
- Cardiovascular models, Regeneration, Time-lapse imaging, Tissue culture
- MeSH Terms
-
- Animals
- Animals, Genetically Modified
- Cell Proliferation
- Heart/physiology*
- Microscopy, Fluorescence
- Organ Culture Techniques/methods*
- Pericardium/cytology*
- Pericardium/physiology*
- Pericardium/transplantation
- Regeneration
- Zebrafish*/genetics
- PubMed
- 27055096 Full text @ Nat. Protoc.
Citation
Cao, J., Poss, K.D. (2016) Explant culture of adult zebrafish hearts for epicardial regeneration studies. Nature Protocols. 11:872-881.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping