A simple technique for isolating healthy heart cells from mouse models.
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.
Shioya
Takao
T
Department of Physiology, Faculty of Medicine, Saga University, Saga 849-8501, Japan. shioya@cc.saga-u.ac.jp
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
D000318:Adrenergic beta-Agonists; D007552:Isotonic Solutions; C037692:Tyrode's solution; D007545:Isoproterenol