Key result
Modified Langendorff perfusion yields ~135 viable mouse SAN myocytes per heart by precisely controlling enzymatic digestion.
Why the study?
Existing methods of isolating sinoatrial node myocytes by direct tissue immersion make it difficult to judge the degree of digestion, cell yield, and viability.
Answers are grounded in this paper and cite their sources.
A modified Langendorff perfusion method provides a more reliable and efficient way to isolate viable sinoatrial node myocytes from adult mice for downstream experiments.
Abstract Sinoatrial node (SAN) is the pacemaker of the heart in charge of initiating spontaneous electronical activity and controlling heart rate. Myocytes from SAN can generate spontaneous rhythmic action potentials, which propagate through the myocardium thereby triggering cardiac myocyte contraction. Acutely, the method for isolating sinoatrial node myocytes (SAMs) is critical in studying the protein expression and function of myocytes in SAN. Currently, the SAMs were isolated by transferring SAN tissue directly into the digestion solution, but it is difficult to judge the degree of digestion, and the cell yield and viability. Here, we present a modified protocol for isolation of SAMs in mice, based on the collagenase II and protease perfusion of heart using a Langendorff-apparatus, and subsequent dissociation of SAMs. The appearance and droplet flow rate of the heart could be significantly changed during enzymatic digestion via perfusion, which allowed us to easily judge the degree of digestion and avoid incomplete or excessive digestion. The SAMs with stable yield and viability achieving from our optimized approach would facilitate the follow-up experiments.
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Li et al. (2023) studied Healthy mice (sinoatrial node myocyte isolation). Langendorff perfusion system for enzymatic digestion vs. Traditional direct tissue digestion was evaluated on Yield of viable sinoatrial node myocytes (SAMs). A modified Langendorff perfusion method for isolating adult mouse sinoatrial node myocytes yielded 120-150 viable cells per heart by allowing precise control of enzymatic digestion.
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