文献类型：期刊文献

中文题名：不同降温速率对脐血干细胞冷冻复苏后生物学特性的影响

英文题名：Effects of Different Cooling Rates on Cryopreservation of Hematopoietic Stem Cells from Cord Blood

作者：沈华萍[1];丁春梅[1];迟占有[1];康自珍[1];谭文松[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,上海200237

年份：2003

卷号：19

期号：4

起止页码：489

中文期刊名：生物工程学报

外文期刊名：Chinese Journal of Biotechnology

收录：MEDLINE(收录号：15969071);CSTPCD;;Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；PubMed;

基金：国家"8 63"计划生物领域项目;上海市现代生物与医药项目基金资助 (No.10 2 12 0 8 0 1;No .0 0 43190 0 3)~~

语种：中文

中文关键词：降温速率;脐血干细胞;冷冻复苏;生物学特性;脐血库

外文关键词：umbilical cord blood bank, hematopoietic cells, cryopreservation

摘要：考察了不同降温速率对脐血造血干细胞各种生物学特性的影响。在 4℃～ - 40℃的降温范围内 ,分别选择 - 0 5℃ min ,- 1℃ min ,- 5℃ min的降温速率进行降温 ,对复苏后的脐血单个核细胞的回收率、活性和CD3 4+含量的变化以及BFU E、CFU GM和CFU MK集落的回收率进行了考察 ,发现在 - 1℃ min的降温速率下 ,脐血MNC回收率可达 93 3 %± 1 8% ,活性可达 95 0 %± 3 9% ,CD3 4+ 细胞回收率达 80 0 %± 17 9% ,BFU E回收率为87 1%± 5 5 % ,CFU GM回收率达 88 5 %± 8 9% ,CFU MK的回收率也达到 86 2 %± 7 4%。并且对复苏后的细胞进一步进行体外培养 ,发现在 - 1℃ min的降温速率下复苏的细胞仍然具有与未经冷冻细胞相似的扩增能力 ,而- 0 5℃ min和 - 5℃ min这两种降温速率条件下复苏的细胞与未经冷冻的细胞相比差距较大。因而 - 1℃
Clinical evidence of hematopoietic restoration with umbilical cord blood (UCB) grafts indicates the UCB can be a useful source of hematopoietic stem cells for routine bone marrow reconstitution. Considering (10±5)×10 8 nucleared cells per cord blood unit, there is a potential limitation for the use of cord blood in adults, which, however, can be overcome by ex vivo expansion of cells. A prerequisite for expansion is the significantly higher recovery of MNC, CD34 +cells and colony-forming cells(CFC) by thawing cryopreserved MNC. Cooling rate always acts as a critical factor that can affect the recovery of cells. Although the rate of -1℃/min is adopted in most of the cryopreservations, no data has been reported about the detailed effects of different cooling rates. The aim of the study was to reveal the different effects of cooling rates on cryopreservation of hematopoietic stem cells from cord blood. UCB samples were collected, and cryopreserved as mononuclear cells (MNC) with different cooling rates of -0 5℃/min, -1℃/min, -5℃/min, and the recovery and viability of MNC and CD34 + cells, the clonogenic capa- city and the ex vivo expansion potential of UCB progenitor cells were evaluated after thawing. With -1℃/min cooling rate, the recovery of MNC reached 93 3%±1 8%, viability 95 0%±3 9%, recovery of CD34 +cells 80 0%±17 9%, and clonogenic recovery were 87 1%±5 5%, 88 5%±8 9%, 86 2%±7 4% for BFU-E、CFU-GM、CFU-MK, respectively. After 14 days of liquid culture, no significant difference was detected in CFC expansion between fresh and cryopreserved MNC cells with -1℃/min cooling rate, but this was not the case with -0 5℃/min and -5℃/min. In conclusion, it was demonstrated that controlling the rate at -1℃/min is more suitable for cryopreservation of hematopoietic stem cells than -0 5℃/min and -5℃/min.