摘要
目的比较两步和三步法分离外周血单个核细胞(PBMC)和血浆的效果。方法对送至样本资源库的16例抗凝血标本分别采用两步法和三步法分离PBMC及血浆,并对2种不同处理方法的分离效果进行比较。两步法为将抗凝血加入淋巴细胞分离液,离心后分别分离PBMC和血浆;三步法则先离心分离血浆后再加入淋巴细胞分离液,离心后分离PBMC。结果 2种处理方法处理后检测的活细胞数、总细胞数及活细胞比例结果相近,差异均无统计学意义(P>0.05)。两步法血浆中细胞因子水平均低于三步法,其中25种细胞因子比较,差异均有统计学意义(P<0.05)。结论两步法省时,操作少,在分离PBMC方面效果与三步法类似;但两步法在分离血浆方面会影响血浆细胞因子水平。
Objective The quality of the specimens affects the subsequent scientific research,and the different separate methods significantly influence the specimen quality.We compared the effects of two different separate methods on the peripheral blood mononuclear cells(PBMC)and plasma.Methods 16 specimens of Biobank were separated respectively by two-step method and three-step method,and then compared the effect of the two different methods.Two-step method:add the anticoagulant blood specimen into the lymphocyte separation medium,then separate the peripheral blood mononuclear cells and plasma after centrifugation.Three-step method:first,separate plasma after centrifugation and dilute the rest specimen;then add the rest specimen into the lymphocyte separation medium;finally,separate PBMC by the second centrifugation.Results To separate PBMC:live cells,total cells and cell viability were similar between two-step method and three-step method,there were no statistical differences.To separate plasma:27 plasma cytokines were analysed,the level of plasma cytokines with two-step method were lower than that of three-step method,25 cytokines were statistically significant difference(P<0.05).Conclusion Compared with three-step method,two-step method has similar effect of separating PBMC with less time and less operation,but the two-step method could reduce the level of plasma cytokines.
作者
丛佳
徐杰
代艳超
王子康
孙坚萍
CONG Jia;XU Jie;DAI Yanchao;WANG Zikang;SUN Jianping(Department of Hematology,Beijing Tongren Hospital,Capital Medical University,Beijing 100730,China;Beijing You'an Hospital,Capital Medical University,Beijing 100069,China)
出处
《国际检验医学杂志》
CAS
2019年第1期40-42,47,共4页
International Journal of Laboratory Medicine
基金
国家传染病重大专项(2017ZX10202102-003-003)
北京市重点实验室项目(BZ0373)
关键词
品种资源
生物标本库
单核细胞
细胞因子
血浆
variety resources
biological specimen banks
monocytes
cytokines
plasma
