Three Cardiomyocyte Isolation Protocols
In single-cell adult cardiomyocyte experiments, isolation is the most critical but oftentimes most difficult part, and a poor isolation can ruin an experiment. There is not yet a consensus on the best isolation method, and this will vary from lab to lab, but there are three different isolation techniques that we recommend trying.
The Langendorff method, in which the aorta is cannulated and retrograde-perfused with enzymatic solutions, is the most widely-used isolation technique. There are two variations on Langendorff isolation that labs have had success with. The first, described by Kaestner et al, uses liberase for digestion, while Liao and Jain prefer collagenase. In their cardiomyocyte isolation review, Louch et al discuss some of the variability and different options of Langendorff isolation, including enzyme selection and whether to use BDM.
Finally, Ackers-Johnson et al have recently presented a novel, non-Langendorff technique that aims to avoid some of the potential pitfalls of Langendorff isolation, like sterility issues, downstream effects of heparin injection, and difficulty mounting the aorta. This new method involves buffer perfusion by direct injection, pH adjustment, and finally aortic clamping to induce deep myocardium perfusion, all within a laminar flow cabinet.
So now that you have a cardiomyocyte isolation protocol in hand, do you know what you need to do next? Check out the IonOptix Application Notes page to learn about loading your cells with Fura-2 AM, acquiring calcium and contractility data, culture pacing procedures, and more!
As always if you have any questions about your IonOptix systems in general, we have a team of scientists and engineers ready to lend you support. Feel free to give us a call directly or inquire at firstname.lastname@example.org.