Trehalose, an easy, safe and efficient cryoprotectant for the parasitic protozoan Trypanosoma brucei.
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.
Center for Parasitic Organisms, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, PR China.
Yan-Zi
YZ
Wen
Center for Parasitic Organisms, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, PR China.
Bi-Xiu
BX
Su
School of Chemistry and Chemical Engineering, Sun Yat-Sen University, Guangzhou 510275, PR China.
Shu-Shen
SS
Lyu
Ecosystems & Environment Centre, School of Environment and Life Sciences, University of Salford, Salford M5 4WT, UK; Biomedical Research Centre, School of Environment and Life Sciences, University of Salford, Salford M5 4WT, UK.
Geoff
G
Hide
Center for Parasitic Organisms, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, PR China.
Zhao-Rong
ZR
Lun
Center for Parasitic Organisms, State Key Laboratory of Biocontrol, School of Life Sciences, Sun Yat-Sen University, Guangzhou 510275, PR China. Electronic address: laidehua@mail.sysu.edu.cn.
De-Hua
DH
Lai
D000818:Animals; D002470:Cell Survival / Q000187:drug effects; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D005615:Freezing; D051379:Mice; D014199:Trehalose / Q000494:pharmacology*; D014346:Trypanosoma brucei brucei / Q000187:drug effects*
D003451:Cryoprotective Agents; D014199:Trehalose
Cryopreservation; Cryoprotective agent; Trehalose; Trypanosoma brucei