Research of the mammalian gut microbiome has in recent years been principally defined by the culture independent analysis of the genetic potential inherent to these microbial communities. However, there has been a renewed interest in culture-based studies of the gut microbiome to increase both the breadth and depth of gut microbial isolates, as these are widely recognised to provide the clearest opportunity to link biological activities with specific microbes. This chapter provides an overview of the methodological approaches widely used to culture fastidious gut microbes, and our evolving understanding of how macro- and micronutrients impact their growth. In addition, alternative culturing approaches are discussed that take advantage of the existing wealth of metagenomic data, including genomic, genetic and antibody-based isolation strategies. Many of these approaches have already been validated and they may enable the development of new efficient high-throughput culturing strategies to aid the recovery of the uncultured microbial majority.