A rapid and efficient method for the isolation of postnatal murine cardiac myocyte and fibroblast cells

作者：Weldrick Jonathan J; Abdul Ghani Mohammad; Megeney Lynn A; Burgon Patrick G^*

来源：Canadian Journal of Physiology and Pharmacology, 2018, 96(5): 535-539.

DOI：10.1139/cjpp-2017-0742

• cardiomyocyte
• isolation
• cardiac
• perfusion
• neonatal
• fibroblast

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.

• 出版日期2018-5