摘要
目的:探索离体造血干细胞(hematopoietic stem cell,HSC)活力维持的相关措施,为超远距离转运非血缘HSC提供技术支持。方法:外周血造血干细胞(peripheral blood hematopoietic stem cell,PBHSC)采集物按不同分组要求给予相应处理后,于4℃冰箱保存,并分别于0、24、48和72 h检测CD34+细胞比例、相对细胞活性、相对细胞增殖率、相对集落形成率、氧分压以及细胞内活性氧(reactive oxygen species,ROS),另设未经处理的PBHSC作为正常对照组。结果:CD34+细胞比例在72 h内未检测到变化。对照组随着保存时间的延长,相对细胞活性、相对细胞增殖率和相对集落形成率均逐渐下降(r=-0. 796、r=-0. 883和r=-0. 815)。血浆稀释、抗氧化剂和充氧均能在一定程度上提高相对细胞活性和相对细胞增殖率,但充氧可降低相对集落形成率。2种或3种因素联合显示出更强的保护作用。细胞内ROS水平随着保存时间的延长而逐渐下降,充氧在提高氧分压的同时,也增加细胞内ROS水平,加入抗氧化剂可降低ROS水平。结论:CD34+细胞百分比不能作为细胞活力观察指标。血浆稀释、充氧和抗氧化剂可提高PBHSC存活和活力,但充氧可增加细胞内ROS水平,不利于PBHSC集落形成能力维持。多因素联合可更好地维持细胞活力。
Objective: To explore the maintaining measures for the vitality of hematopoietic stem cells ( HSC) in vitro,so as provide technical support for ultra long distance transport of HSC collected from unrelated donors.Methods: Peripheral blood hematopoietic stem cells ( PBHSC) were treated by different methods according to various groups,then stored at 4 ℃ in the refrigerator.The percentage of CD34^+ cells,relative cell activity,relative cell proliferation rate,relative colony - forming rate,oxygen fraction and intracellular reactive oxygen species ( ROS) were detected at 0,24,48 and 72 h after storage of PBHSC respectively.Results: The percentage of CD34^+ cells during 72 h storage did not altered.Along with the prolonging of storage time,the relative cell activity,relative cell proliferation rate and relative colony - forming rate gradually decreased in untreated PBHSC( control group) ,the related coefficients were - 0.796,- 0.883 and - 0.815 respectively.Plasma dilution,antioxidants and oxygenation could improve the relative cell activity and relative cell proliferation rate,but oxygenation could decrease the relative colony - forming rate of PBHSC.The combination of 2 or 3 factors showed stronger protection effects on PBHSC.The intracellular level of ROS decreased gradually with the prolonging of storage time.Oxygenation of PBHSC could increase oxygen fraction,and also increase the intracellular level of ROS at the same time.The addition of antioxidants could reduce the level of ROS.Conclusion:The percentage of CD34^+ cells can not serve as the indicator of PBHSC vitality.Plasma dilution,oxygenation and antioxidants can increase the survival and viability of PBHSC,but oxygenation can increase the intracellular ROS level and impair colony - forming ability of PBHSC.The combination of multiple factors can maintain the vitality of PBHSC better.
作者
刘佳欣
马巍娜
谭奕
梁永清
尹文杰
岑坚
黄友章
沈建良
LIU Jia-Xin;MA Wei-Na;TAN Yi;LIANG Yong-Qing;YIN Wen-Jie;CEN Jian;HUANG You-Zhang;SHEN Jian-Liang(Department of Hematology,Navy General Hospital of Chinese People's Liberation Army,Beijing 100048,China;Beijing Marrow Donor Program,Beijing 100073,China)
出处
《中国实验血液学杂志》
CAS
CSCD
北大核心
2018年第6期1772-1779,共8页
Journal of Experimental Hematology
基金
北京市财政项目PXM2015-077203-000004
关键词
造血干细胞
细胞活力
细胞活力维持
hematopoietic stem cell
cell vitality
cell vitality maintenance
