2种细胞冻存液对脐血单个核细胞冻存质量的影响  

The influence of two different cryopreservation solution on mononuclear cells from cord blood

作　　者：黄馨萍 林科佳 魏宗科 魏志璋 罗晓玲 王宇环 

机构地区：[1]深圳市合一康生物科技股份有限公司技术研发中心,深圳518045

出　　处：《广西医科大学学报》2017年第9期1270-1273,共4页Journal of Guangxi Medical University

基　　金：广东省科技计划项目(No.2014B09091036)

摘　　要：目的:探讨2种细胞冻存液对脐血来源单个核细胞冻存质量的影响,为脐血来源单个核细胞的冻存提供合适的冻存方法和理论依据。方法:采用细胞计数仪检测不同冻存液(冻存液1:培养基∶自体血浆∶DMSO=5∶4∶1;冻存液2:自体血浆∶DMSO=9∶1)对脐血单个核细胞复苏后回收总数的影响;同时使用流式细胞仪和体外造血细胞培养术分析脐血细胞损失的类群和脐血造血分化的能力。结果:细胞计数仪和台盼蓝拒染实验发现,冻存液2细胞总回收率为(72.88±12.24)%,显著高于冻存液1的细胞总回收率(56.65±9.34)%;流式结果也表明,冻存液1的CD34+造血干细胞损失率高达(35.90±2.56)%,其他CD14+单核/中性粒细胞、CD4+T细胞、CD8+T细胞和CD3-CD56+NK细胞损失率分别为(49.95±3.68)%、(24.21±2.35)%、(33.87±3.21)%和(35.18±3.52)%;而冻存液2的细胞除了CD4+T细胞和CD8+T细胞损失率分别为(13.50±1.25)%和(24.67±3.72)%,其他细胞比例都相应增加。同时,集落形成能力检测结果显示,冻存液2的CFU-GM和BFU-E的集落团多于冻存液1的细胞。结论:自体血浆∶DMSO=9∶1的冻存液对脐血单个核细胞冻存效果较好。Objective： To investigate the effect of two cryopreservation solution on mononuclear cells of cord blood and provide ideal cryopreservation solution based on reasonable theories. Methods： The total number of cord blood cells cryopreserved by two cryopreservation solution （Group 1： culture medium ： autologous plasma ： DMSO=5：4：1 ： Group 2： autologous plasma ： DMSO=9：1 ）were counted by cell counting instrument. Meanwhile, the cell loss rate of cord blood cells and hematopoietic differentiation ability of cord blood cells were respective- ly assessed by FACS and MethodCult assay. Results： The results from the cell count and trypan blue staining ex- periments showed the total cellular recovery of cryopreservation group 2 was （72.88± 12.24）%, which was sig- nificantly higher than that of cryopreservation group 1 （ 56. 65±9. 34） %; flow cytometry data also indicated that the CD34＋cell loss rate was as high as （35.90±2.56）%, and other cell loss rates like CD14＋ monocytes neutro- phils, CD4＋ T, CD8＋ T and CD3- CD56~ NK cell was （49.95 ± 3.68）%, （24. 21 ± 2. 35）%, （33.87± 3.21 ）% and（35. 18± 3. 52）%, respectively. Whereas the proportion of other cells in cryopreservation group 2 was in- creased except CD4＋ T and CD8＋T cell loss which was （13. 50± 1.25）% and （24. 67± 3.72）%. Even the re- suits of hematopoietic differentiation ability showed that the CFU-GM and BFU-E colony groups of cryopreser- vation group 2 were also significantly more than those of cryopreservation group 1. Conclusion： The cryo- preservation effect of cryopreservation group 2 （autologous plasma ： DMSO=9：1 ）was better for cryopreservation of umbilical cord blood mononuclear cell.

关 键 词：造血干细胞 脐血 单核细胞 冻存 

分 类 号：R743.33[医药卫生—神经病学与精神病学]