Scientific Publications Database

Article Title: A rapid and efficient method for the isolation of postnatal murine cardiac myocyte and fibroblast cells
Authors: Weldrick, Jonathan J.; Abdul-Ghani, Mohammad; Megeney, Lynn A.; Burgon, Patrick G.
Journal: CANADIAN JOURNAL OF PHYSIOLOGY AND PHARMACOLOGY Volume 96 Issue 5
Date of Publication:2018
Abstract:
The capacity to isolate and study single cardiomyocytes has dramatically enhanced our understanding of the fundamental mechanisms of the heart. Currently, 2 primary methods for the isolation of cardiomyocytes are employed: (i) the neonatal isolation protocol and (ii) the Langendorff isolation method. A major limiting feature of both procedures is the inability to isolate cardiomyocytes between 3 days and 3 weeks after birth. Herein, we report the establishment and validation of a new method for the rapid and efficient isolation of mouse cardiomyocytes, regardless of age. This novel procedure utilizes whole heart perfusion of a trypsin-collagenase Krebs-based buffer through the left ventricle at a high flow rate. Cardiomyocytes can be isolated in significantly less time with a simple, syringe-pump-based apparatus. Typically, we can digest 10-15 hearts per hour. Altogether, we have established an efficient and reproducible method for the rapid isolation of fresh cardiomyocytes from postnatal mouse hearts of any age.