The present report describes a method for establishment of colonies of Blastocystis hominis from single cells in soft agar. The percentage of colony-forming efficiency (% CFE = number of colonies grown/number of cells inoculated x 100) for the cultures was greatly improved by the addition of sodium thioglycollate. Five human Blastocystis isolates chosen for this study showed no apparent variation in colonial morphology. Isolated colonies were also successfully grown in liquid medium, providing a means of obtaining large numbers of B. hominis cells that had arisen from a single clone.