In vitro culture of epicardial cells from adult zebrafish heart on a fibrin matrix
- Authors
- Kim, J., Rubin, N., Huang, Y., Tuan, T.L., and Lien, C.L.
- ID
- ZDB-PUB-120125-27
- Date
- 2012
- Source
- Nature Protocols 7(2): 247-255 (Journal)
- Registered Authors
- Lien, Ching-Ling (Ellen)
- Keywords
- none
- MeSH Terms
-
- Animals
- Cell Culture Techniques*
- Cell Movement
- Cell Proliferation
- Culture Media
- Epithelial-Mesenchymal Transition
- Fibrin
- Heart/physiology
- Pericardium/cytology*
- Regeneration
- Tissue Culture Techniques
- Zebrafish*
- PubMed
- 22262006 Full text @ Nat. Protoc.
We describe here a protocol for culturing epicardial cells from adult zebrafish hearts, which have a unique regenerative capacity after injury. Briefly, zebrafish hearts first undergo ventricular amputation or sham operation. Next, the hearts are excised and explanted onto fibrin gels prepared in advance in a multiwell tissue culture plate. The procedure allows the epicardial cells to outgrow from the ventricle onto a fibrin matrix in vitro. This protocol differs from those used in other organisms by using a fibrin gel to mimic blood clots that normally form after injury and that are essential for proper cell migration. The culture procedure can be accomplished within 5 h; epicardial cells can be obtained within 24–48 h and can be maintained in culture for 5–6 d. This protocol can be used to investigate the mechanisms underlying epicardial cell migration, proliferation and epithelial-to-mesenchymal transition during heart regeneration, homeostatic cardiac growth or other physiological processes.