<p>Fungal lipids have garnered significant attention due to their environmentally friendly properties. However, their commercialization remains limited, partly because of the lack of robust fermentation strategies, suitable microbial strains, and low lipid productivity. Moreover, only a small number of microbial lipids have been evaluated at the pilot scale, highlighting the need for further studies to assess their technological feasibility and environmental benefits. Therefore, high cell density cultivation (HCDC) is essential for enhancing microbial lipid accumulation and improving product yields. This strategy enables the generation of elevated biomass levels in well-defined fermentation media, thereby facilitating the production of substantial amounts of lipids. Multiple biotechnological approaches, including medium optimization, modulation of environmental and nutritional factors, and genetic engineering, can enhance lipid production in various oleaginous microorganisms, although these improvements remain limited by the intrinsic metabolic capacities of each species. This review highlights various HCDC strategies, including the oleaginous fungi and yeasts commonly employed, the range of available feedstocks, and the environmental and nutritional parameters that influence cell growth and lipid accumulation. It also examines recent advances in genetic tools and metabolic engineering approaches aimed at improving microbial lipid production through targeted gene modification. This review also outlines recent advances in lipid biosynthesis pathways and the mechanisms underlying lipid accumulation. In addition, it summarizes the applications and economic value of microbially derived lipids across various industrial sectors. Overall, integrating diverse fermentation strategies with emerging technological innovations has the potential to significantly enhance microbial lipid production, thereby increasing their biotechnological and nutraceutical relevance.</p> Graphical abstract <p></p>