影响人外周血单个核细胞4℃保存效果的因素分析

Factors Affecting the Preservation of Human Peripheral Blood Mononuclear Cells at 4℃

目的:分析4℃无血清条件下人外周血单个核细胞保存效果及其相关影响因素。方法:采用密度梯度离心法分离人外周血单个核细胞,使用不同细胞密度、添加人血清白蛋白、葡萄糖等条件对其进行4℃保存,并检测保存72 h内细胞的存活率、细胞数量、活细胞数量和细胞表型。结果:随保存时间延长,人外周血单个核细胞数量逐步下降(r=0.982),与(5-6)×10^(6)/ml细胞密度相比较,细胞保存密度为(1-2)×10^(6)/ml时细胞数量下降趋势减缓;添加人血清白蛋白和葡萄糖可有效提升人外周血单个核细胞的存活率,其中,2%人血清白蛋白保存效果更优;淋巴细胞亚群的分析结果表明,相较于对照组,添加人血清白蛋白和葡萄糖后NK细胞和T细胞数量下降趋势明显减缓。结论:(1-2)×10^(6)/ml的细胞密度、2%人血清白蛋白更适合PBMC的保存,5%葡萄糖可提升人外周血单个核细胞的4℃保存效果。

Objective:To analyze the preservation effect and related influencing factors of human peripheral blood mononuclear cells under serum-free condition at 4℃.Methods:Human peripheral blood mononuclear cells were isolated by density gradient centrifugation,and stored at 4℃under different cell concentrations,supplemented with human serum albumin,and glucose.The cell viability,total cell number,viable cell number and cell phenotype were detected during preservation of 72 h.Results:With the prolongation of storage time,the number of human peripheral blood mononuclear cells gradually decreased(r=0.982).Compared with the cell concentration of(5-6)×10^(6)cells/ml,the cell number decreased more slowly when the cell storage concentration was(1-2)×10^(6)cells/ml;Adding human serum albumin and glucose can effectively improve the survival rate of human peripheral blood mononuclear cells,among which 2%human serum albumin has a better preservation effect;Compared with the blank control group,the analysis results of cell subsets showed that the downward trends of NK cells and T cells were significantly slowed after adding albumin and glucose.Conclusion:The cell density of(1-2)×10^(6)/ml and 2%human serum albumin are more suitable for the preservation of PBMC,and 5%glucose can improve the preservation effect of human peripheral blood mononuclear cells at 4℃.