3D tumoroids have revolutionized in vitro/ex vivo cancer biology by recapitulating the complex diversity of tumors. While tumoroid provide new insights into cancer development and treatment response, several limitations remain. The tumor microenvironment, especially the immune system, strongly influences tumor development. Absence of immune cells in tumoroids may yield flawed conclusions. Macrophages, key players in tumor progression, pose integration challenges into tumoroid. In this study, we established three optimized and standardized methods for co-culturing human macrophages with breast cancer tumoroids: a semi-liquid model and two matrix-embedded models tailored for specific applications. Optimizing culture conditions, we tracked interactions and macrophage infiltration using flow cytometry and light sheet microscopy. Macrophages not only influenced tumoroid molecular profiles but also chemotherapy response. This underscores the importance of increasing the complexity of current 3D models to more accurately reflect in vivo conditions. The three models have different application and can be used in various ways.