Evaluation of trehalose and sucrose as cryoprotectants for hematopoietic stem cells of umbilical cord blood
Evaluation of trehalose and sucrose as cryoprotectants for hematopoietic stem cells of umbilical cord blood
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DOI：
10.1016/j.cryobiol.2008.01.003
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发表时间：
2008-04-01
期刊：
影响因子：
2.7
通讯作者：
Porto, L. C.
中科院分区：
文献类型：
--
作者：
Rodrigues, J. P.;Paraguassu-Braga, F. H.;Porto, L. C.
骨髓移植(BMT)是一种涉及造血干细胞(HSC)移植的治疗程序。到目前为止，临床上使用的造血干细胞有三种来源：骨髓、动员的外周血和脐带血。根据干细胞来源或移植类型的不同，这些细胞会被冷冻保存。最广泛使用的冷冻保护剂是10%(v/v)的二甲基亚砜(Me2SO)，但输注Me2SO-冷冻保存的细胞经常与患者的严重副作用有关。在这项研究中，我们评估了海藻糖和蔗糖在减少Me2SO用量的情况下用于脐血细胞冷冻保存的效果。复苏后的细胞用台盼蓝计数和活性检测，用流式细胞仪检测HSC比例，用集落形成单位(CFU)法测定造血祖细胞比例。在冻存至少2周后，30 mmol/L海藻糖+2.5%Me2SO(v/v)或60 mmol/L蔗糖+5%Me2SO(v/v)的冻存脐血，其祖细胞克隆形成潜能、细胞活力和CD45(+)/34(+)细胞数与10%(v/v)Me2SO相似。因此，脐带血和其他HSC一样，可以在添加二糖的情况下，用标准Me2SO浓度的四分之一进行冷冻保存。在低温冻存液中使用低浓度的Me2SO可以减少对患者的副作用，从而提高造血细胞移植的安全性。(C)2008 Elsevier Inc.保留所有权利。
Bone marrow transplantation (BMT) is a therapeutic procedure that involves transplantation of hematopoietic stem cells (HSC). To date, there are three sources of HSC for clinical use: bone marrow; mobilized peripheral blood; and umbilical cord blood (UCB). Depending on the stem cell source or type of transplantation, these cells are cryopreserved. The most widely used cryoprotectant is dimethylsulfoxide (Me2SO) 10% (v/v), but infusion of Me2SO-cryopreserved cells is frequently associated with serious side effects in patients. In this study, we assessed the use of trehalose and sucrose for cryopreservation of UCB cells in combination with reduced amounts of Me2SO. The post-thawed cells were counted and tested for viability with Trypan blue, the proportion of HSC was determined by flow cytometry, and the proportion of hematopoeitic progenitor cells was measured by a colony-forming unit (CFU) assay. A solution of 30 mmol/L trehalose with 2.5% Me2SO (v/v) or 60 mmol/L sucrose with 5% Me2SO (v/v) produced results similar to those for 10% (v/v) Me2SO in terms of the clonogenic potential of progenitor cells, cell viability, and numbers of CD45(+)/34(+) cells in post-thawed cord blood cryopreserved for a minimum of 2 weeks. Thus, cord blood, as other HSC, can be cryopreserved with 1/4 the standard Me2SO concentration with the addition of disaccharides. The use of Me2SO at low concentrations in the cryopreservation solution may improve the safety of hematopoietic cell transplantation by reducing the side effects on the patient. (C) 2008 Elsevier Inc. All rights reserved.