The information gained from the Human Genome Project has facilitated molecular as well as cellular studies not only to find the origins of Breast Cancer (BC), but also to create novel, and effective treatments. In order to provide an infrastructure for local and international research in this area, Iranian Center for Breast Cancer (ICBC) has established a Bio-Bank (BB) for BC. This article describes the aim, structure, and activities in general, and the challenging issues confronting the bank as a model for the establishment of Bio-Banks in developing countries in particular. The methods employed by the Bank could be explained in the following categories: Blood and Tissue sampling, Preparation and Banking of collected Samples, Clinical and Histopathology data collection, Collaboration Protocol, Challenging issues, and the programs to confront the problems. During the five-year activity of the bank, 110 families were enrolled for genetic counseling, from whom 600 biologic samples were obtained, including 387 blood samples and 213 tissue samples. Of 387 blood samples, 317 (82%) were found to belong to the BC patients and the remaining 70 (18%) belonged to their available relatives. The number of samples increased over the study period partly as a result of the programs designed to confront the problems. During the study period, there were some finished research studies using the samples of BB, and many other studies which are still ongoing. ICBC-BB is a model of biologic sample banking which provides a significant number of biological samples for local and international collaborative research projects regarding molecular and cellular aspects of BC. In establishing the ICBC-BB we have experienced problems and challenges, some general and some local. Some were expected and others not, but we have identified solutions.