title: Isolation and culture of adult mouse cardiomyocytes for cell signaling and in vitro cardiac hypertrophy.
abstract: Technological advances have made genetically modified mice, including transgenic and gene knockout mice, an essential tool in many research fields. Adult cardiomyocytes are widely accepted as a good model for cardiac cellular physiology and pathophysiology, as well as for pharmaceutical intervention. Genetically modified mice preclude the need for complicated cardiomyocyte infection processes to generate the desired genotype, which are inefficient due to cardiomyocytes' terminal differentiation. Isolation and culture of high quantity and quality functional cardiomyocytes will dramatically benefit cardiovascular research and provide an important tool for cell signaling transduction research and drug development. Here, we describe a well-established method for isolation of adult mouse cardiomyocytes that can be implemented with little training. The mouse heart is excised and cannulated to an isolated heart system, then perfused with a calcium-free and high potassium buffer followed by type II collagenase digestion in Langendorff retrograde perfusion mode. This protocol yields a consistent result for the collection of functional adult mouse cardiomyocytes from a variety of genetically modified mice.
authors.affiliation: Department of Biochemistry and Cancer Biology, University of Toledo College of Medicine and Life Sciences.
authors.forename: Daxiang
authors.initials: D
authors.lastname: Li
authors.affiliation: Department of Biochemistry and Cancer Biology, University of Toledo College of Medicine and Life Sciences.
authors.forename: Jian
authors.initials: J
authors.lastname: Wu
authors.affiliation: Department of Biochemistry and Cancer Biology, University of Toledo College of Medicine and Life Sciences.
authors.forename: Yan
authors.initials: Y
authors.lastname: Bai
authors.affiliation: Department of Physiology and Pharmacology, University of Toledo College of Medicine and Life Sciences.
authors.forename: Xiaochen
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Department of Biochemistry and Cancer Biology, University of Toledo College of Medicine and Life Sciences; lijun.liu@utoledo.edu.
authors.forename: Lijun
authors.initials: L
authors.lastname: Liu
mesh_terms: D000818:Animals; D006332:Cardiomegaly / Q000473:pathology*; D002450:Cell Communication / Q000502:physiology*; D003584:Cytological Techniques / Q000379:methods*; D051379:Mice; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000473:pathology