Dendritic cells (DCs) are highly potent antigen-presenting cells crucial for the innate and adaptive immune response and for maintaining immune tolerance towards self-antigens. Although they share many common features, multiple DC subtypes with different immune functions have been identified. Originally, DCs were considered to be cells with purely myeloid origin. Recent studies have now demonstrated that DCs can also develop from lymphatic progenitors. Various cytokines and transcription factors are known to be responsible for the development of DC subpopulation. Depending on the subpopulation and the maturation state of these cells, they are either able to induce a broad cytotoxic immune response, and therefore represent a promising tool for anticancer vaccination therapies in humans or induce immune tolerance and are important within the context of autoimmunity. This review will focus on recent advances on the identification of different DC subpopulations including phenotypical and functional differences and on recent developments on protocols for IN VITRO generation of myeloid-derived DCs.