Three-dimensional (3D) cell cultures have long been recognized as a tool for the study of tissue architecture, polarity, and invasion. However, only recently these systems have been used to study centrosome and cilia functions. Studying these organelles in 3D cultures has elucidated new functions that otherwise would have been overlooked, demonstrating the value of these experimental systems to the field. Here we describe a culture method to study mammary epithelial cells in a 3D environment.