The developmental anatomy of human cardiac outflow was studied in a series of 16 normal embryos (gestational days 29-39, crown-rump length 6-20 mm, stages 14-19). Structural features and kinetics during truncal septation (TS) were described from external photographs, serial histological sections, and computer graphic reconstructions of selected tissues. Early in the period studied, the tubular myocardium ensheathed the single cardiac lumen and spiralling conotruncal ridges, which were filled with mesenchymal cells during days 31-33. As TS began (late stage 16), the aorticopulmonary (AP) septum appeared across the dorsal wall of the aortic sac between arches IV and VI. Mesenchymal condensations formed within the AP septum, crossing the lumen bifurcation to extend along the truncal ridges to the myocardium. During days 35-37, the cephalic margin of the myocardium grew or folded in toward these mesenchymal condensations between the developing valves and within the nearby conal ridges, which appeared to fuse to separate the subvalvular outflow channels by day 39. These observations are consistent with studies in chicks and rats which suggest that mesenchymal condensations or cell death foci interact with the distal myocardial rim during TS to form a structural septation complex dividing the two arterial streams.