Ex vivo bone organ cultures preserve cell-cell and cell-matrix interactions and allow the study of cancer and bone cells in their natural tumor environment. This chapter is a comprehensive guide detailing the methods employed to establish and characterize ex vivo bone-cancer organ cultures. The chapter begins by outlining the materials and step-by-step procedures for successfully initiating and maintaining these cultures, emphasizing the advantage of modeling cancer within the bone microenvironment. Additionally, the chapter provides a detailed account of the techniques utilized to analyze key endpoints, such as tumor growth, microenvironmental changes, and therapeutic responses.