Development and Evaluation of Modified Cryopreservation for Long-Term Storage of Blastocystis Subtypes 1–3 and 6



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 3 months, 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.

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This article has been extracted from the thesis written by Mr. Seyed Ahmad Karamati in School of Medicine, Shahid Beheshti University of Medical Science (Registration No: M341). The project was also financially supported by Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences (grant No. 887). The authors would like to thank all the staffs of the Foodborne and Waterborne Diseases Research Center for their collaboration.


This study was financially supported by the Research Institute for Gastroenterology and Liver Diseases, Shahid Beheshti University of Medical Sciences, Tehran, Iran (grant no. 887).

Author information




Conceived and designed the experiments: HM SAK MN. Performed the experiments: HM SAK. Analyzed the data: HM SAK. Contributed reagents/materials/analysis tools/positive samples: MN AH HAA MRZ. Wrote the paper: HM SAK SJST. All authors read and approved the final version of the manuscript.

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Correspondence to Maryam Niyyati or Hamed Mirjalali.

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Karamati, S.A., Asadzadeh Aghdaei, H., Niyyati, M. et al. Development and Evaluation of Modified Cryopreservation for Long-Term Storage of Blastocystis Subtypes 1–3 and 6. Acta Parasit. 65, 535–540 (2020).

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  • Blastocystis
  • Subtypes
  • Cryopreservation
  • Long-term storage