| 李 博,王 娜,郑小红,孙可帅,刘雁声,韩 英.利用流式细胞术评价长期冻存对PBMCs各亚型的影响[J].,2020,(18):3413-3418 |
| 利用流式细胞术评价长期冻存对PBMCs各亚型的影响 |
| Using Flow Cytometry to Evaluate the Effects of PBMC Subtypes after Long-term Cryopreservation |
| 投稿时间：2020-03-03 修订日期：2020-03-28 |
| DOI：10.13241/j.cnki.pmb.2020.18.003 |
| 中文关键词: 外周血单个核细胞 低温冻存 淋巴细胞 |
| 英文关键词: PBMCs Cryopreservation Lymphocyte |
| 基金项目:国家自然科学基金项目（81770569）；中国博士后科学基金项目（2018M643871）；国家重大新药创制项目（2014ZX09508002-001） |
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| 中文摘要: |
| 摘要 目的：利用流式细胞术，检测长期冻存后PBMCs总数及各亚型的变化，评价PBMCs的长期冻存效果。方法：收集志愿者外周血PBMCs，利用流式细胞术分析液氮冻存后PBMCs细胞总数及其亚型的变化。结果：长期冻存后，PBMCs总细胞量和细胞活力无显著改变（P=0.19, P=0.32）；T细胞、NK细胞和NKT细胞比例无明显变化，但B细胞比例增多(P<0.01)，单核细胞比例显著减少(P<0.001)；低温保存影响活化的T细胞和Tregs细胞数量（P<0.05, P<0.05），其中初始Tregs和记忆Tregs显著减少（P<0.05, P<0.01）。结论：PBMCs长期冻存会影响B细胞，单核细胞、活化T细胞和Tregs细胞的活性。 |
| 英文摘要: |
| ABSTRACT Objective: To explore the influence of long-term cryopreservation on the total number and subpopulation distribution of PBMCs by flow cytometry, in order to evaluate this storage method. Methods: PBMCs were isolated from the peripheral blood that collected from the volunteers. After long-term cryopreservation in liquid nitrogen, the changes of the total cell number of PBMCs and a serious of surface markers of different subpopulations were analyzed by flow cytometry. Results: The total cell number and survival rate of PBMCs did not significantly change after long-term cryopreservation (P=0.19, P=0.32). Similarly, there was no dramatic alteration in proportion of T cells, NK cells and NKT cells. While the proportion of B cells significantly increased (P<0.01), and the proportion of monocytes cells significantly decreased (P<0.001). Additionally, the proportions of activated T cells and Treg cells were dramatically affected by long-term cryopreservation（P<0.05, P<0.05）. The naive Treg cells and memory Treg cells were significantly decreased (P<0.05, P<0.01). Conclusion: Long-term cryopreservation had a dramatic influence on proportion of B cells, monocytes, activated T cells and Treg cells in PBMCs. |
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