Introduction <p>Diatoms, a diverse group of photosynthetic unicellular algae, have gained significant global attention due to their ecological importance and multifaceted applications in scientific research. Their ecological roles are critical, encompassing nutrient cycling, carbon sequestration, and primary productivity, which establish them as essential components of aquatic food webs. The remarkable species richness of diatoms underscores their evolutionary success and highlights their integral roles in both freshwater and marine ecosystems. In addition to their environmental significance, diatoms possess a rich biochemical profile comprising valuable compounds with immense potential for biotechnological applications. These applications span diverse fields, including biofuel production, pharmaceuticals, and wastewater remediation. Despite the vast diversity and biochemical richness of diatoms, laboratory cultivation and maintenance remain challenging. To address these challenges, two primary methodologies have been developed: xenic and axenic culture techniques. Xenic culture involves maintaining diatoms alongside associated microorganisms, thereby replicating natural conditions and preserving ecological interactions. In contrast, axenic culture techniques focus on isolating pure diatom strains by employing meticulous sterilization processes, enabling precise experimental manipulation and fundamental research. Understanding the significance of xenic and axenic cultivation methodologies is essential for unlocking the full potential of diatoms across diverse scientific domains. This review elaborates on the methodologies, scope, and applications of xenic and axenic culture techniques for diatoms. By examining the intricacies of these cultivation approaches, it seeks to provide insights into optimizing diatom culture practices, advancing research initiatives, and harnessing the biotechnological potential of these extraordinary microorganisms.</p> Clinical trial registration <p>Not applicable.</p>