Representing a fundamental part of the human microbiota, the oral microbial community is characterized by its diverse and unique composition. Oral diseases like dental caries and periodontitis are directly associated with oral microorganisms. Therefore, it is crucial to deepen our understanding of oral microbiota. Advances in high-throughput sequencing technology provide extensive information of the diversity of oral uncultivated microorganisms, prompting an increasing need for microbial isolation and cultivation techniques. This review presents recent research progress on uncultivated oral microorganisms and lists factors that possibly hinder isolation attempts. In addition, advances in methodologies and techniques used for culturing previously uncultured microorganisms and their applications in oral microbiology studies are summarized, giving valuable insights into various aspects of uncultivated oral microorganisms.