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Comparison between two step and three step method to separate the anticoagulant blood specimens of Biobank

CONG Jia;XU Jie;DAI Yanchao;WANG Zikang;SUN Jianping;Department of Hematology,Beijing Tongren Hospital,Capital Medical University;Beijing You′an Hospital,Capital Medical University;  
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(P0.05).Conclusion Compared with three-step method,twostep 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.
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