Cytokinesis, the last major step of cell division, is a complex multistage process involving specific rearrangement of cellular cytoskeleton and a flurry of vesicular transport activities at the cell division plane. Vesicular traffic from the exocytic pathway and the endocytic/recycling pathway, operating again after being shut down since prophase, are engaged to supply the mitotic midzone with materials that would facilitate furrowing, midbody thinning and subsequent abscission of daughter cells. Cytokinesis is spatial and temporally regulated by mitotic kinases, and involves modulation by Arf and Rab small GTPases and their effectors. The latter include vesicle targeting and tethering molecules such as motor proteins, tethering complexes and SNAREs. The process of abscission requires the ultimate engagement of endosomal sorting complex required for transport (ESCRT) complexes. Although a good deal of details remains to be deciphered, cytokinesis in eukaryotes could essentially be visualized as a specialized cellular event requiring complex spatial and temporal regulated processes of membrane traffic.