| The aim of this experiment was to investigate the effects of Pluronic F68 and its combination with DMSO on cryopreservation of cord blood hematopoietic stem cell under -80℃ and provide evidence for clinical usage. The MNCs were separated from cord blood by density gradients of 1.080 Percoll, and were divided into four groups on the basis of different cryopreservation solution which was CP-1, F68 combined with DMSO, F68 alone, and DMSO alone. Each group containing 10 cord blood samples was stored in -80℃ freezer for 90 days. At 30, 60, 90 day, cells were thawed and cell viability was tested by Typan blue exclusion, clonogenic potential and 3H-TdR incorporation. Cell microstructures were also observed under electronic microscope at the end of 90 day. The result showed that F68 alone had cryoprotective effect and the new cryoprotectant containing 5% F68, 5% DMSO and 4% Alb had the best cryoprotective effect compared to the counterparts of 10% DMSO and CP-1. After cryopreservation and thawing, MNC recovery can achieve(83.4±6.3)%,CFU-GM recovery can achieve(68.8±7.9)% and viability by Typan blue exclusion was (92.5±5.1)% at the 90th day. Cell viability had no significant decline within 90 days. Under electronic microscope, less cell damages were observed in group CP-2. The study showed that F68 and its combination with DMSO had ideal protective effects on the cryopreservation of cord blood hematopoietic stem cell.
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