Improvement of the cryopreservation method for the Babesia gibsoni parasite by using commercial freezing media

Kodai Kusakisako; Tatsunori Masatani; Yurika Yada; Melbourne Rio Talactac; Emmanuel Pacia Hernandez; Hiroki Maeda; Masami Mochizuki; Tetsuya Tanaka

doi:10.1016/j.parint.2016.02.012

Improvement of the cryopreservation method for the Babesia gibsoni parasite by using commercial freezing media

Parasitol Int. 2016 Oct;65(5 Pt B):532-535. doi: 10.1016/j.parint.2016.02.012. Epub 2016 Feb 26.

Authors

Kodai Kusakisako¹, Tatsunori Masatani², Yurika Yada³, Melbourne Rio Talactac¹, Emmanuel Pacia Hernandez³, Hiroki Maeda¹, Masami Mochizuki¹, Tetsuya Tanaka⁴

Affiliations

¹ Laboratory of Infectious Diseases, Joint Faculty of Veterinary Medicine, Kagoshima University, Korimoto, Kagoshima 890-0065, Japan; Department of Pathological and Preventive Veterinary Science, The United Graduate School of Veterinary Science, Yamaguchi University, Yoshida, Yamaguchi 753-8515, Japan.
² Transboundary Animal Diseases Research Center, Joint Faculty of Veterinary Medicine, Kagoshima University, Korimoto, Kagoshima 890-0065, Japan.
³ Laboratory of Infectious Diseases, Joint Faculty of Veterinary Medicine, Kagoshima University, Korimoto, Kagoshima 890-0065, Japan.
⁴ Laboratory of Infectious Diseases, Joint Faculty of Veterinary Medicine, Kagoshima University, Korimoto, Kagoshima 890-0065, Japan; Department of Pathological and Preventive Veterinary Science, The United Graduate School of Veterinary Science, Yamaguchi University, Yoshida, Yamaguchi 753-8515, Japan. Electronic address: tetsuya@ms.kagoshima-u.ac.jp.

PMID: 26921519
DOI: 10.1016/j.parint.2016.02.012

Abstract

In vitro cultivation and cryopreservation under liquid nitrogen have already been reported and established for Babesia bovis and Babesia bigemina parasites. Although the in vitro cultivation methods for Babesia gibsoni have been reported and established, the cryopreservation methods for this parasite have not been established completely. In this paper, we compared several freezing media for the cryopreservation of B. gibsoni parasite. The CELLBANKER® series (1 plus and 2), STEM-CELLBANKER®, and CultureSure® were used for commercial freezing media; 10% dimethyl sulfoxide in 90% fetal bovine serum, 20% polyvinylpyrrolidone in phosphate-buffered saline (established for bovine Babesia parasites), and 28% glycerol supplemented with 3% sorbitol and 0.65% NaCl dissolved in water (established for Plasmodium parasites) were used for conventional media. Our results demonstrated that the CELLBANKER® series (1 plus and 2), STEM-CELLBANKER®, and CultureSure® are effective freezing media for B. gibsoni parasite compared to the cryopreservation methods of bovine Babesia and Plasmodium parasites. Our improved method of cryopreservation would contribute to the stability of the in vitro cultivation of B. gibsoni parasite.

Keywords: Babesia gibsoni; Cryopreservation; Freezing medium.