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《Journal of Naval General Hospital》 2006-03
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Cryopreservation of Hemotopoietic Stem Cells Using Dimethyl Suflfoxide and Hydroxyethyl Starch in Low Temperature Refrigerator and Autocontrolled Programmed Cryogenic

HUANG You-zhang~1,SHEN Jian-liang~1,YANG Ping-di~1,XU Shi-xia~2,WU Nan-hai~3,XIANG Dan~1,GONG Li-zhong~1,CEN Jian~1,WANG Li-xin~1,ZHENG Pei-hao~1(1.Department of Hematology,Naval General Hospital,Beijing 100037,China;2.Department of Information,Naval General Hospital,Beijing 100037,China;3.Department of Pediatrics,Naval General Hospital,Beijing 100037,China)  
Objective To explore proper preservatives and simple conditions for cryoprereservation of hemotopoietic stem cells.Methods Bone marrow cells and peripheral blood mononucleated cells obtained by blood cell seperator were added to 5% dimethyl suflfoxide(DMSO) and 6% hydroxyethyl starch(HES,5%DMSO-6%HES),then preserved in low temperature refrigerator.Granulocyte-macrophage colony forming unit(GM-CFU),long term culture initiating cells(LTC-IC),CD34~+ cells and typhan blue resistance cells(TBRC) were counted compared with the cells preserved in 10% DMSO by autocontrolled programmed cryogenic system in liquid nitrogen.Results CFU-GM,LTC-IC,CD34~+ cells and TBRC showed no significant difference between low temperature refrigerator group and liquid nitrogen group.These parameters were not significantly different within one year when the cells were preserved in liquid nitrogen after laid in low tempreture refrigerator,or directly in low tempreture refrigerator,or in liquid nitrogen with autocontrolled programmed cryogenic system,however the cell recovery rate decreased after half a year when the cells were directly preserved in low tempreture refrigerator.The cell activity declined after recovery at room temperature if the preservatives were not removed or diluted in both groups,the group of 10%DMSO was more affected than 5%DMSO-6%HES group was.Conclusion 5%DMSO-6%HES is better than 10%DMSO as preservative for hemopoietic stem cells.The cells in the three groups above will be preserved well within one year.If the cells will be preserved for long duration,liquid nitrogen is the best choice.The preservatives should be diluted immediately after the cells are recovered.
【CateGory Index】: R318.52
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