Exocrine, endocrine, and neuroendocrine cells store hormones and neuropeptides in secretory granules (SGs), which undergo regulated exocytosis in response to an appropriate stimulus. These cargo proteins are sorted at the trans-Golgi network into forming immature secretory granules (ISGs). ISGs undergo maturation while they are transported to and within the F-actin-rich cortex. This process includes homotypic fusion of ISGs, acidification of their lumen, processing, and aggregation of cargo proteins as well as removal of excess membrane and missorted cargo. The resulting mature secretory granules (MSGs) are stored in the F-actin-rich cell cortex, perhaps as segregated pools exhibiting specific responses to stimuli for regulated exocytosis. During the last decade our understanding of the maturation of ISGs advanced substantially. The use of biochemical approaches led to the identification of membrane molecules mechanistically involved in this process. Furthermore, live cell imaging in combination with fluorescently tagged marker proteins of SGs provided insights into the dynamics of maturing ISGs, and the functional implications of cytoskeletal elements and motor proteins.