To evaluate the effects of gestational age on left and right ventricular diastolic filling in human fetuses, we retrospectively reviewed the diastolic flow velocity patterns through mitral and tricuspid valves in 307 normal fetuses aged 17 to 39 weeks' gestation. The subjects were divided into 3 age groups; 17 to 24 weeks, 25 to 31 weeks, and 32 to 39 weeks. The variables measured were peak flow velocities of early diastole (peak E wave), of atrial contraction (peak A wave), and the velocity ratio (peak E/A ratio). As a whole, the transmitral peak E wave and peak E/A ratio correlated with age using a second-order polynomial curve fit. The strength of the linear correlation between age and transmitral peak E wave and peak E/A ratio and the slope of the relation were greater in the group aged 32 to 39 weeks than in group aged 25 to 31 weeks. Similar temporal change was observed in the relation between age, transtricuspid peak E wave, and peak E/A ratio. The peak A wave for both atrioventricular valves showed little change with gestational age. Contrary to the accepted concept that fetal peak E wave and peak E/A ratio increases linearly with aging, this study shows that early diastolic filling increases mainly after 25 weeks' gestation. We speculate that the maturational changes in ventricular properties in human fetuses accelerate after midgestation.