Blastocystis is a protozoan parasite living in the intestine of humans and a wide range of animals. Although Blastocystis grows in several cultivation media, axenification and serial cultivations for long time are the main challenges of the researchers. Therefore, the long-term storage of subtypes/strains of Blastocystis using cryopreservation provides a suitable source of this parasite for the physiological, biochemical, and biological studies. In the current study, seven xenic isolates including two separated isolates from ST1-3 and one isolate from ST6 were cryopreserved using a standard method with minor modifications. After 3months, all isolates were recovered and cultivated in DMEM medium. The findings of the method showed all seven isolates were successfully recovered in DMEM medium. In addition, all isolates remained viable after several sub-cultures. It seems that cryopreservation is a simple method that can provide a suitable condition for the long-term storage of Blastocystis.