To find the key factors that were involved in the survival and vascular endothelial differentiation of chick blatodisc induced by fibroblast growth factor 2 (FGF-2), we built a chick vasculogenesis model in vitro. Subsequently, the activities of phosphatidylcholine-specific phospholipase C (PC-PLC), including Ca(2+)-dependent and -independent PC-PLC, and the level of reactive oxygen species (ROS) were evaluated during the endothelial differentiation of chick blastodisc. The results showed that Ca(2+)-indepentent PC-PLC underwent a remarkable increase in 24 h (P < 0.01), then it decreased gradually with the cell differentiation, while the Ca(2+)-depentent PC-PLC was nearly not changed in the whole process. At the same time, ROS level dramatically decreased during the cell differentiation. To understand the role of PC-PLC and how it performs its function in the vascular endothelial differentiation induced by FGF-2, we suppressed PC-PLC activity by its specific inhibitor D609 (tricyclodecan-9-yl potassium xanthate) at 24 h during the cell differentiation. As a result, the cell differentiation could not progress and the intracellular level of ROS was elevated. The data suggested that PC-PLC and ROS were involved in chicken blastodisc differentiation to vascular endothelial cells. PC-PLC was an important factor in the blastodisc cell survival and differentiation, and it might perform its function associated with ROS.