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Microplastic pollution caused by washing processes of synthetic textiles has recently been assessed as the main source of primary microplastics in the oceans. Therefore, understanding the effective contribution of the washing process of synthetic clothes to this environmental problem, is of great importance. In this study, wash trials at real scale were performed on commercial clothes by using a household washing machine in order to gain reliable data about the release of microplastics, and to identify possible influences of textile characteristics on the release. The wastewater was collected and filtered through subsequent filters with decreasing porosity, and the amount and dimensions of microfibres were determined. Microfibre release was analysed in relation to the nature and characteristics of the washed clothes. Results showed that microfibres released during washing range from 124 to 308 mg for kg of washed fabric depending from the type of washed garment that corresponds to a number of microfibres ranging from 640,000 to 1,500,000. Some textile characteristics, such as the type of fibres constituting the yarns and their twist, influenced the release of microfibres during washing. A great amount of microfibres of cellulosic nature was also released during washing of clothes made with a blend of polyester/cellulose. Finally the most abundant fraction of microfibres shed was retained by filters with pore size of 60 µm, presenting an average length of 360–660 μm and an average diameter of 12–16 μm, indicating dimensions that could pass through wastewater treatment plants and pose a threat for marine organisms.