从滤白后的白细胞滤器中回收外周血单个核细胞的可行性研究  

The feasibility of PBMCs recovery from white cell-reduced filters after filtration

作　　者：陈小林[1] 李冬妹[1] 徐彬 贾延军[1] 王明慧[1] Chen Xiaolin;Li Dongmei;Xu Bin;Jia Yanjun;Wang Minghui(Blood Components Section,Beijing Red Cross Blood Center,Beijing 100088,China)

机构地区：[1]北京市红十字血液中心成分科,100088

出　　处：《北京医学》2020年第4期340-343,共4页Beijing Medical Journal

摘　　要：目的研究从滤白后的白细胞滤器中回收外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)的可行性。方法分别于全血过滤完成后1 h和18 h进行PBMCs回收。使用生理盐水反向冲洗使用过的白细胞滤器,分离PBMCs,部分用于流式细胞术(fluorescence activated cell sorting,FACS)分析,部分用于冻存,并分别于冻存后1周、1个月、3个月和6个月通过FACS分析其中各种有核细胞的含量和活性。结果反向冲洗后1 h内处理,可从过滤400 ml全血的白细胞滤器中分离得到(1.85±0.84)×10^8个PBMCs,其中淋巴细胞占大多数,其次为单核细胞,而大部分粒细胞都被去除[淋巴细胞占(73.58±13.12)%,单核细胞占(14.91±7.14)%,粒细胞占(11.49±10.21)%]。FACS检测结果显示,PBMCs中粒细胞、单核细胞和淋巴细胞的活性与白膜层中分离出的PBMCs比较,差异无统计学意义(P>0.05)。除粒细胞外,细胞冻存后6个月内其他有核细胞,如单核细胞和淋巴细胞的比例和活性与冻存前相比差异无统计学意义。结论利用滤白后的白细胞滤器回收PBMCs是可行的,本研究为更充分地利用白细胞滤器提供了实验数据支持。Objective To verify the feasibility of peripheral blood mononuclear cells(PBMCs)recovery from white cell-reduced filters after filtration.Methods The PBMCs was recovered from white cell-reduced filters at one hour and 18 hours after the whole blood filtration.The used white cell-reduced filters were reversely flushed with saline and the PBMCs were isolated.Part of them were used for flow cytometry analysis and others were used for cryopreservation.The content and activity of various nucleated cells in PBMCs were analyzed by flow cytometry.Results If performed in one hour after reverse flushing,(1.85±0.84)×10^8 PBMCs could be isolated from the white cell-reduced filters of 400 ml whole blood,with lymphocytes accounting for the majority[(73.58±13.12)%],followed by monocytes[(14.91±7.14)%],but most of the granulocytes were removed[(11.49±10.21)%].The results of flow cytometry test showed that there was little difference between the activity of granulocytes,monocytes and lymphocytes in PBMCs and that of those cells in PBMCs isolated from the buffy coats.In addition to granulocytes,there was no significant difference in the proportion and activity of other nucleated cells,such as monocytes and lymphocytes,when comparing these cells cryopreserved within six months with before cryopreservation.Conclusions It is feasible to recycle PBMCs from white cell-reduced filters.The results of this study provide data support for making fuller use of the white cell-reduced filters.

关 键 词：外周血单个核细胞 白细胞滤器 流式细胞术 

分 类 号：R457.1[医药卫生—治疗学]