AbstractThe proepicardium (PE) is an embryonic progenitor cell population, which provides the epicardium, the majority of the cardiac interstitium, the coronary vasculature and possibly some cardiomyocytes. Recent studies have documented (1) the presence of bilaterally paired PE anlagen in several vertebrates, and (2) species‐specific differences in the fate of the left and right PE anlagen. Here, we document PE development in Xenopus laevis (stages 37–46). The PE appears at stage 41 in the form of a cone‐shaped accumulation of mesothelial cells covering the pericardial surface of the right horn of the sinus venosus. No such structure appears on the left sinus horn. At the end of stage 41, the tip of the PE establishes a firm contact with the developing ventricle. A secondary tissue bridge is established facilitating the transfer of PE cells to the heart. During stages 41–46, this tissue bridge is visible in vivo through the transparent body wall. Corresponding to the morphological data, the PE marker gene Tbx18 is expressed only on the right sinus horn suggesting a right‐sided origin of the PE. Left–right lineage tracing has confirmed this idea. These results show that Xenopus PE development proceeds in a bilaterally asymmetric pattern as previously observed in chicks. We speculate that asymmetric PE development is controlled by signals from left–right signaling pathways and that the PE is an indicator for right‐sidedness in Xenopus embryos. Xenopus might be a good model to uncover the role of left–right signaling pathways in the control of asymmetric PE development. Developmental Dynamics 237:3088–3096, 2008. © 2008 Wiley‐Liss, Inc.