Microbial communities in marine habitats are regarded as underexplored reservoirs for discovering new natural products with potential application. However, only a few microbes in nature can be cultivated in the laboratory. This has led to the development of a variety of isolation and cultivation methods, and in situ cultivation is one of the most popular. Diverse in situ cultivation methods, with the same basic principle, have been applied to a variety of environmental samples. Compared with conventional approaches, these new methods are able to cultivate previously uncultured and phylogenetically novel microbes, many with biotechnological potential. This review introduces the various in situ cultivation methods for the isolation of previously uncultured microbial species and their potential for marine microbial resource mining. Furthermore, studies that investigated the key and previously unidentified mechanisms of growing uncultivated microorganisms by in situ cultivation, which will shed light on the understanding of microbial uncultivability, were also reviewed.