Tompa Andrea, Johansson Junko, Islander Ulrika, Faresjö Maria
Department of Clinical Diagnostics, School of Health and Welfare, Jönköping University, SE-551 11 Jönköping, Sweden.
Division of Systems and Synthetic Biology, Department of Life Sciences, Chalmers University of Technology, SE-412 96 Gothenburg, Sweden.
Biomedicines. 2024 Oct 11;12(10):2319. doi: 10.3390/biomedicines12102319.
: Flow cytometry is a convenient tool in immunophenotyping for monitoring the status of immunological conditions and diseases. The aim of this study was to investigate the effect of isolation and cryopreservation by flow cytometric analysis on subpopulations of CD4 T helper (Th), T regulatory (Treg), CD8 T cytotoxic (Tc), CD56 NK, CD19 B and monocytes. Freshly isolated and cryopreserved peripheral blood mononuclear cells (PBMCs) were compared to fresh whole blood. : Peripheral blood was collected from healthy donors and prepared for flow cytometric analysis using the same panels of antibodies throughout the study. : Comparisons between fresh (F)- and cryopreserved (C)-PBMCs showed no major differences in percentages of CD4, Th1, Th2 and CD4CD25CD127 Treg cells. No differences in percentage of CD8 or subpopulations of naive/stem, central or effector memory cells were observed between F- and C-PBMCs. The percentage of CD56 NK cells, CD19 B cells or classical and nonclassical monocytes did not differ between F-and C-PBMCs either. On the contrary, whole blood had lower percentages of Th and NK cells but higher percentages of Th1, Th17, Th1Th17, Tregs, Tc and B cells compared to C-PBMCs, while it had a higher proportion of Tc compared to F-PBMCs. : Flow cytometric immunophenotyping minimally differs between freshly isolated and cryopreserved PBMCs. This implies the possibility of cryostorage of cohorts for later analysis. Importantly, care must be taken when comparing results from whole blood with isolated and cryopreserved PBMCs. Collectively, these results can contribute to the standardization of flow cytometric protocols in both clinical and research settings.
流式细胞术是免疫表型分析中用于监测免疫状态和疾病的便捷工具。本研究旨在通过流式细胞术分析研究分离和冷冻保存对CD4辅助性T细胞（Th）、调节性T细胞（Treg）、CD8细胞毒性T细胞（Tc）、CD56自然杀伤细胞（NK）、CD19 B细胞和单核细胞亚群的影响。将新鲜分离和冷冻保存的外周血单个核细胞（PBMC）与新鲜全血进行比较。：从健康供体采集外周血，并在整个研究过程中使用相同的抗体组合进行流式细胞术分析准备。：新鲜（F）和冷冻保存（C）的PBMC之间的比较显示，CD4、Th1、Th2和CD4CD25CD127 Treg细胞的百分比没有重大差异。F和C PBMC之间未观察到CD8或幼稚/干细胞、中枢或效应记忆细胞亚群百分比的差异。F和C PBMC之间的CD56 NK细胞、CD19 B细胞或经典和非经典单核细胞的百分比也没有差异。相反，与C PBMC相比，全血中Th和NK细胞的百分比更低，但Th1、Th17、Th1Th17、Treg、Tc和B细胞的百分比更高，而与F PBMC相比，全血中Tc的比例更高。：新鲜分离和冷冻保存的PBMC之间的流式细胞术免疫表型分析差异极小。这意味着可以对队列进行冷冻保存以供后续分析。重要的是，在将全血结果与分离和冷冻保存的PBMC结果进行比较时必须谨慎。总体而言，这些结果有助于临床和研究环境中流式细胞术方案的标准化。
