The paper presents the processes of entrepreneurial human capital accumulation and its impact on rural business growth. Data are derived from four surveys on rural businesses in mountainous and less favoured areas in Southern Europe. Formal pathways of entrepreneurial human capital accumulation refer to education and training, while informal pathways include the cognitive processes of work and managerial experience acquisition and the non-cognitive processes of being raised within an entrepreneurial family environment and/or being raised in the area within which the business is later set-up. The studies reveal that there is a variety of processes of entrepreneurial human capital and knowledge accumulation that are case study specific. Human capital accumulation processes related to education and training or to work and managerial experience still plays the prime role in predicting successful businesses. Results indicate the need for decentralised, flexible and selective entrepreneurial human capital accumulation support programmes that take into account local idiosyncrasies and needs.