The genus Cephaleuros comprises plant parasitic algae distributed in tropical and subtropical zones globally. This study aimed to select culture media and optimize the culture conditions for Cephaleuros species. An extensive survey was conducted in southern Thailand and five species were found, namely Cephaleuros diffusus on baegu leaves, C. expansa on orange jasmine leaves, C. karstenii on cacao leaves, C. pilosa on mangosteen leaves, and C. virescens on soursop leaves. To select suitable culture media, the five Cephaleuros species were cultured on Bold’s basal medium (BBM), bristol medium, high salt medium (HSM), trebouxia medium, trebouxia bristol medium, and modified BBM. All the Cephaleuros species grew well in BBM, followed by HSM and bristol media. Bold’s basal medium amended with indole- 3-acetic acid (IAA) was found to be the most suitable for growth of all these Cephaleuros species when compared with plain BBM. Furthermore, culture under 24 h light maximized growth of the five Cephaleuros species. The results showed that BBM amended with IAA was the most suitable medium for Cephaleuros species cultures under incubation with 24 h light. Gametangia-like body and sporangia were formed on BBM after 3 mo of incubation. Further research is needed regarding cultures of pure Cephaleuros species on synthetic media in genetic studies and in tests of pathogenicity in plants.