title: In Vivo Cannulation Methods for Cardiomyocytes Isolation from Heart Disease Models.
abstract: Isolation of high quality cardiomyocytes is critically important for achieving successful experiments in many cellular and molecular cardiology studies. Methods for isolating cardiomyocytes from the murine heart generally are time-sensitive and experience-dependent, and often fail to produce high quality cells. Major technical difficulties can be related to the surgical procedures needed to explant the heart and to cannulate the vessel to mount onto the Langendorff system before in vitro reperfusion can begin. During this period, transient hypoxia and ischemia may damage the heart, resulting in low yield and poor quality of cells, especially for heart disease models that have fragile cells. We have developed novel in vivo cannulation methods to minimize hypoxia and ischemia, and fine-tuned the entire protocol to produce high quality ventricular myocytes. The high cell quality has been confirmed using important structural and functional criteria such as morphology, t-tubule structure, action potential morphology, Ca2+ signaling, responsiveness to beta-adrenergic agonist, and ability to have robust contraction under mechanically loaded condition. Together these assessments show the preservation of the cardiac excitation-contraction machinery in cells isolated using this technique. The in vivo cannulation method enables consistent isolation of high-quality cardiomyocytes, even from heart disease models that were notoriously difficult for cell isolation using traditional methods.
authors: Department of Pharmacology, University of California Davis, Davis, California, United States of America.
authors: Zhong
authors: http://orcid.org/0000-0003-1072-5788
authors: Z
authors: Jian
authors: Department of Pharmacology, University of California Davis, Davis, California, United States of America.|Microsurgery Core of Department of Pharmacology, University of California Davis, Davis, California, United States of America.
authors: Yi-Je
authors: YJ
authors: Chen
authors: Department of Pharmacology, University of California Davis, Davis, California, United States of America.|Department of Biomedical Engineering, University of California Davis, Davis, California, United States of America.
authors: Rafael
authors: R
authors: Shimkunas
authors: Department of Pharmacology, University of California Davis, Davis, California, United States of America.
authors: Yuwen
authors: Y
authors: Jian
authors: Department of Pharmacology, University of California Davis, Davis, California, United States of America.
authors: Mark
authors: M
authors: Jaradeh
authors: Department of Pharmacology, University of California Davis, Davis, California, United States of America.
authors: Karen
authors: K
authors: Chavez
authors: Department of Internal Medicine, Division of Cardiology, University of California Davis, Davis, California, United States of America.
authors: Nipavan
authors: N
authors: Chiamvimonvat
authors: Department of Medicine and Cellular & Molecular Medicine, University of Arizona, Tucson, Arizona, United States of America.
authors: Jil C
authors: JC
authors: Tardiff
authors: Department of Pharmacology, University of California Davis, Davis, California, United States of America.
authors: Leighton T
authors: LT
authors: Izu
authors: Veterans Administration Healthcare System, Cardiology Section, University of California San Diego, San Diego, California, United States of America.
authors: Robert S
authors: RS
authors: Ross
authors: Department of Pharmacology, University of California Davis, Davis, California, United States of America.|Department of Internal Medicine, Division of Cardiology, University of California Davis, Davis, California, United States of America.|Department of Biomedical Engineering, University of California Davis, Davis, California, United States of America.
authors: Ye
authors: Y
authors: Chen-Izu
mesh_terms: D000200:Action Potentials / Q000502:physiology; D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000818:Animals; D001011:Aorta; D020013:Calcium Signaling; D006328:Cardiac Catheterization / Q000295:instrumentation / Q000379:methods*; D002339:Carotid Arteries; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D007545:Isoproterenol / Q000494:pharmacology; D008297:Male; D008810:Mice, Inbred C57BL; D008822:Mice, Transgenic; D009200:Myocardial Contraction; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*
chemical_list: D000318:Adrenergic beta-Agonists; D007545:Isoproterenol