Abstract:

BACKGROUND: As the classical cryoprotectant for hematopoietic stem cells, dimethylsulfoxide has considerably toxicity for both the thawed cells and the patients. Trehalose is a stable and unharmful disaccharide, which has been widely used in the field of the cryopreseved red cells, platelet and organs.
OBJECTIVE: To explore the feasibility of trehalose used as cryoprotectant for hematopoietic stem cells.
METHODS: After recombinant colony stimulating factor mobilization, mononuclear cells were separated from hematopoietc stem cells and were divided into the control, 0.5 mol/L trehalose and 1.0 mol/L trehalose groups. The hematopoietc stem cells were auto-controlled programmed cryogenic system, then stored in liquid nitrogen for 1 week, followed by rapidly thawed in a 40 ℃ water bath. Cell survival rates were detected by trypan blue exclusion assay; the recovery rates of colony-forming unit granulocyte-macrophage and CD34⁺ cells were counted by CD34-PE/CD45-FITC double-staining methods and flow cytometry.
RESULTS AND CONCLUSION: Compared with the control group, cell survival, colony-forming unit granulocyte-macrophage and CD34⁺ cells recovery rate of the 0.5 and 1.0 mol/L trehalose groups were obviously increased, especially in the 0.5 mol/L trehalose group (P < 0.001 or P < 0.01). Trehalose has some cryoprotective potential for cryopreservation of hematopoietic stem cells, particularly, 0.5 mol/L exerts a superior cryoprotective potential.