Trehalose, an easy, safe and efficient cryoprotectant for the parasitic protozoan Trypanosoma brucei

Wen, YZ; Su, BX; Lyu, SS; Hide, G; Lun, ZR; Lai, DH

Abstract

Trehalose, a non-permeating cryoprotective agent (CPA), has been documented as less toxic and highly efficient at cryopreserving different kinds of cells or organisms. In the present study, trehalose was evaluated for its application in cryopreservation of both Trypanosoma brucei procyclic and bloodstream form cells. The cryopreservation efficiency was determined by the motility of trypanosomes after thawing, as well as a subsequent recovery and infectivity assessment. The viability of trypanosomes from cultivation that were frozen in a serial concentrations of trehalose showed similar results to classical CPAs of glycerol and DMSO. Nevertheless, trypanosomes cryopreserved in 0.2M trehalose showed the best growth characteristic during subsequent cultivation. In addition, CPA cocktails with trehalose and permeating CPA glycerol or DMSO were developed and evaluated. Interestingly, trypanosomes in host (mouse) blood cryopreserved in 0.4M trehalose plus 5% glycerol showed higher infectivity than those preserved in trehalose/DMSO cocktails as well as individually. Further investigations showed that, in comparison with slow freezing at -80°C, flash freezing in liquid nitrogen provided better cryopreservation for bloodstream form cells than slow freezing. In conclusion, trehalose is an easy, safe and efficient CPA for cryopreservation of T. brucei and potentially for other protozoan species and cells

Citation

Wen, Y., Su, B., Lyu, S., Hide, G., Lun, Z., & Lai, D. (2016). Trehalose, an easy, safe and efficient cryoprotectant for the parasitic protozoan Trypanosoma brucei. Acta tropica, 164, 297-302. https://doi.org/10.1016/j.actatropica.2016.09.024

• Journal Article Type: Article
• Acceptance Date: Sep 25, 2016
• Online Publication Date: Sep 27, 2016
• Publication Date: Dec 1, 2016
• Deposit Date: Oct 13, 2016
• Publicly Available Date: Oct 13, 2016
• Journal: Acta Tropica
• Print ISSN: 0001-706X
• Publisher: Elsevier
• Volume: 164
• Pages: 297-302
• DOI: https://doi.org/10.1016/j.actatropica.2016.09.024
• Publisher URL: http://dx.doi.org/10.1016/j.actatropica.2016.09.024
• Related Public URLs: http://ac.els-cdn.com/S0001706X16305502/1-s2.0-S0001706X16305502-main.pdf?_tid=a8d1767c-9146-11e6-aa41-00000aacb35d&acdnat=1476364514_96007479def4c69ddf622aa839298cc0; http://www.salford.ac.uk/environment-life-sciences/els-academics/geoff-hide
• Additional Information: Funders : National Sciences Foundation of China;Fundamental Research Funds for the Central Universities; Projects : Trehalose, an easy, safe and efficient cryoprotectant for the parasitic protozoan Trypanosoma brucei; Grant Number: #31272305, #31472058, #31301876, #31402029; Grant Number: #141gpy23