title: In vitro culture of epicardial cells from adult zebrafish heart on a fibrin matrix.
abstract: 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.
authors.affiliation: The Saban Research Institute of Children's Hospital Los Angeles, Los Angeles, California, USA.
authors.forename: Jieun
authors.initials: J
authors.lastname: Kim
authors.forename: Nicole
authors.initials: N
authors.lastname: Rubin
authors.forename: Ying
authors.initials: Y
authors.lastname: Huang
authors.forename: Tai-Lan
authors.initials: TL
authors.lastname: Tuan
authors.forename: Ching-Ling
authors.initials: CL
authors.lastname: Lien
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D002465:Cell Movement; D049109:Cell Proliferation; D003470:Culture Media; D058750:Epithelial-Mesenchymal Transition; D005337:Fibrin; D006321:Heart / Q000502:physiology; D010496:Pericardium / Q000166:cytology*; D012038:Regeneration; D046509:Tissue Culture Techniques; D015027:Zebrafish
chemical_list: D003470:Culture Media; D005337:Fibrin