Adult mice were anesthetized with 3% isoflurane and injected with 200 U of heparin 5 min before heart excision. The heart was quickly cannulated and retrogradely exposed to perfusion buffer (120.4 mM NaCl, 14.7 mM KCl, 0.6 mM KH2PO4, 0.6 mM Na2HPO4, 1.2 mM MgSO4, 10 mM Hepes, 4.6 mM NaHCO3, 30 mM taurine, 10 mM BDM (2,3-butanedione monoxime), and 5.5 mM glucose) at 37°C. Hearts were then perfused for 12 min with enzyme solution containing 50 mg of collagenase and 0.005% trypsin in perfusion buffer containing 12.5 μM CaCl2. Solution was then switched to perfusion buffer for another 3 min. Atria was removed, and ventricles were gently minced. Cardiomyocytes were dissociated by pipetting in stop solution (perfusion buffer containing 10% fetal bovine serum and 12.5 μM CaCl2). Cells were serially exposed to increasing amounts of calcium (62 μM to 1 mM).

Note: The content above has been extracted from a research article, so it may not display correctly.

Please log in to submit your questions online.
Your question will be posted on the Bio-101 website. We will send your questions to the authors of this protocol and Bio-protocol community members who are experienced with this method. you will be informed using the email address associated with your Bio-protocol account.

We use cookies on this site to enhance your user experience. By using our website, you are agreeing to allow the storage of cookies on your computer.