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Annually, researchers from around the world, who work in the field of mammary gland biology and breast cancer and related areas, have a great opportunity to meet and discuss their work at a conference organized by the European Network of Breast Development and Cancer (ENBDC). These meetings, entitled Annual ENBDC Workshop: Methods in mammary gland biology and breast cancer, are largely methodologically oriented. They enable not only presentation of the latest scientific results, but also dissemination of cutting-edge approaches and forefront technologies that have facilitated these discoveries. The latest meeting took place on the 16th to 18th of May 2019 in Weggis, Switzerland, and presented exciting findings achieved using high resolution ‘omics approaches, genetic mouse models, organoids, and state-of-the-art imaging. Here, we present a collection of articles based on or related to the topics of the ENBDC workshop.