Cell swelling is a major problem of cardiac preservation. This study evaluated the effect of storage solution osmolality on long-term preservation of cardiac function. Isolated rat hearts were flushed and stored in solutions of 260 to 350 mOsm/kg water at 0°C for 9 hr. Cardiac performance was assessed using the isolated working heart reperfusion. Function of fresh unstored hearts served as the control. The stored hearts had normal heart rate after reperfusion. Other hemodynamic functions recovered to various levels of the control value and showed biphasic responses to solution osmolality. Hearts stored in hyposmotic (260 and 270 mOsm) and hyperosmotic (310, 330, and 350 mOsm) solutions performed poorly. Those stored in 280 to 290 mOsm solutions showed superior recovery in cardiac function. Among all parameters, coronary flow correlated linearly with aortic flow, cardiac output, systolic pressure, work, oxygen consumption, and coronary vascular resistance. Solutions of 280, 290, and 310 mOsm caused 6 to 9% increases in tissue water (TW) over the control hearts during 9 hr storage, whereas 350 mOsm solution maintained TW at the control level. After reperfusion, all stored hearts gained water (25 to 38%) compared to hearts with no reperfusion. Postreperfusion myocardial ATP content was only 70-79% of control level and did not correlated to function recovery. In conclusion, (1) the optimal osmolality of our preservation fluid is 280 to 290 mOsm and moderate or severe hyperosmolality is detrimental to cardiac function, and (2) preservation of coronary function may be crucial to future improvement of storage solution. © 1990.