title: A simple technique for isolating healthy heart cells from mouse models.
abstract: Single heart cells of mouse models provide powerful tools for heart research. However, their isolation is not easy, and it imposes a significant bottleneck on their use in cellular studies of the heart. Aiming to overcome this problem, this report introduces a novel technique that reproducibly isolates healthy heart cells from mouse models. Using simple devices that ensure easy handling and the rapid aortic cannulation of a small mouse heart, cell isolation was done under physiological conditions without using the "KB" medium or 2,3-butanedione monoxime (BDM). The isolated cells consistently had a healthy appearance and a high viability of 75 +/- 5% (mean +/- SD) in Tyrode solution containing 1.8 mM Ca2+. After 8 h of storage at 37 degrees C, they still had a viability of 45 +/- 12%. The cells showed normal contraction properties when field-stimulated, and they generated normal action potentials and membrane currents under the whole-cell clamp condition. The beta-adrenergic signal transduction of the cells was also normal when it was examined with the isoproterenol enhancement of the L-type Ca2+ current.
authors.affiliation: Department of Physiology, Faculty of Medicine, Saga University, Saga 849-8501, Japan. shioya@cc.saga-u.ac.jp
authors.forename: Takao
authors.initials: T
authors.lastname: Shioya
mesh_terms: D000200:Action Potentials / Q000502:physiology; D000318:Adrenergic beta-Agonists / Q000494:pharmacology; D000818:Animals; D002404:Catheterization / Q000295:instrumentation; D002469:Cell Separation / Q000379:methods*; D002470:Cell Survival; D006262:Health; D007545:Isoproterenol / Q000494:pharmacology; D007552:Isotonic Solutions; D051379:Mice; D008810:Mice, Inbred C57BL; D023421:Models, Animal; D009200:Myocardial Contraction / Q000502:physiology; D032383:Myocytes, Cardiac / Q000166:cytology* / Q000187:drug effects / Q000502:physiology*; D018408:Patch-Clamp Techniques; D013997:Time Factors
chemical_list: D000318:Adrenergic beta-Agonists; D007552:Isotonic Solutions; C037692:Tyrode's solution; D007545:Isoproterenol