抄録
Single heart cells from genetically-engineered mouse models are of great importance for studying physiology and pathophysiology of the heart. With single-cell measurement techniques such as patch-clamp and calcium imaging, they provide powerful tools for unveiling molecular mechanisms underlying heart functions. Doing single-cell experiments with these techniques demand a consistent supply of quality heart cells; however, it is not easy to isolate such quality cells from mouse models. This has long been a major problem for utilizing mouse models, obstructing their use in heart research. With the aim to overcome this problem, we developed a simple method for isolating single heart cells from mouse models. By the use of simple tools to facilitate handling of their tiny hearts and a refined protocol to minimize their ischemic damages, this method isolates single heart cells of mouse models with great ease and consistency. Cell isolation is done under physiological condition, without using "KB" medium or BDM that has long been used for relieving cell damages. Thus isolated heart cells are rod-shaped, quiescent, and relaxed in Tyrode solution containing 1.8 mM Ca, while they have a consistent 75 +/- 5% (mean +/- SD, n = 5) viability under this condition. Lifetime of these cells is long enough for doing same-day experiments, since they retain 45 +/- 12% viability after being stored for 8 hours in Tyrode solution at 37'C. The cells have normal morphology, electrophysiological and contraction properties. They are expected to facilitate the use of genetically-engineered mouse models in heart research. [J Physiol Sci. 2007;57 Suppl:S202]
