Abstract: The assembly of a bipolar spindle is crucial for symmetric partitioning of duplicated chromosomes during cell division. Centrosomes (spindle pole body [SPB] in yeast) constitute the two poles of this bipolar structure and serve as microtubule nucleation centers. A eukaryotic cell enters the division cycle with one centrosome and duplicates it before spindle formation. A proteinaceous link keeps duplicated centrosomes together until it is severed at onset of mitosis, enabling centrosomes to migrate away from each other and assemble a characteristic mitotic spindle. Hence, centrosome separation is crucial in assembly of a bipolar spindle. Whereas centrosome (or SPB) duplication has been characterized in some detail, the separation process is less well understood. Here, we review recent studies that uncover new players and provide a greater understanding of the regulation of centrosome (or SPB) separation.