[1]户乃丽,徐晓雪,邹林樾,等.FACSAriaⅢ流式细胞仪分选小鼠肺脏自然杀伤细胞方法的建立与评价[J].医学信息,2021,34(15):115-118.[doi:10.3969/j.issn.1006-1959.2021.15.030]
　HU Nai-li,XU Xiao-xue,ZOU Lin-yue,et al.Establishment and Evaluation of A Method for Sorting Mouse Lung Natural Killer Cells by FACSAria Ⅲ Flow Cytometer[J].Medical Information,2021,34(15):115-118.[doi:10.3969/j.issn.1006-1959.2021.15.030]
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FACSAriaⅢ流式细胞仪分选小鼠肺脏自然杀伤细胞方法的建立与评价()
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医学信息[ISSN:1006-1959/CN:61-1278/R]

卷:
34卷
期数:
2021年15期
页码:
115-118
栏目:
论著
出版日期:
2021-08-01

文章信息/Info

Title:
Establishment and Evaluation of A Method for Sorting Mouse Lung Natural Killer Cells by FACSAria Ⅲ Flow Cytometer
文章编号:
1006-1959（2021）15-0115-04
作者:
户乃丽; 徐晓雪; 邹林樾; 等
（首都医科大学中心实验室，北京 100069）
Author(s):
HU Nai-li; XU Xiao-xue; ZOU Lin-yue; et al.
(Central Laboratory,Capital Medical University,Beijing 100069,China)
关键词:
流式细胞; NK细胞; 细胞存活率
Keywords:
Flow cytometry; NK cells; Cell survival rate
分类号:
R-331
DOI:
10.3969/j.issn.1006-1959.2021.15.030
文献标志码:
A
摘要:
目的 利用流式细胞仪分选小鼠肺脏自然杀伤细胞（NK细胞）并检测分选后细胞纯度与存活率。方法 小鼠肺脏经胶原酶灌注法得到单细胞悬液，流式细胞仪分选悬液中FVS510-CD3-CD19-NK1.1+NKp46+细胞，分选后的细胞流式细胞仪回测阳性率，台盼蓝染色检测细胞活力。结果 第一滴液流断点位置Drop1为345，Gap为9，液流开启0.5 h以上无晃动，无断点位置改变，稳定性良好，各种参数符合分选条件。分选前NK细胞占总细胞（4.40±1.20）% Total，分选后为（96.10±2.20）% Total，差异有统计学意义（P＜0.05）。经台盼蓝染色显示，分选后细胞存活率为（95.70±2.02）%，细胞形态良好。结论 利用流式细胞分选仪可快速高效分离小鼠肺脏NK细胞，分选后的细胞保持良好的活性，可为进一步研究NK细胞提供保证。
Abstract:
Objective To sort natural killer cells (NK cells) from mouse lungs by flow cytometry and to detect the purity and survival rate of the cells after sorting.Methods Mouse lungs were perfused with collagenase to obtain a single cell suspension, and FVS510-CD3-CD19-NK1.1+NKp46+ cells in the suspension were sorted by flow cytometry.After sorting, flow cytometry back-tested the positive rate of cells, and trypan blue staining to detect cell viability.Results The breakpoint position of the first drop flow was Drop1 was 345, Gap was 9, and the flow was turned on for more than 0.5 h without shaking, no breakpoint position change, good stability, and various parameters were in line with the sorting conditions.Before sorting, NK cells accounted for (4.4±1.2)% Total,After sorting, it was (96.10±2.20)% Total, the difference was statistically significant (P<0.05).The trypan blue staining showed that the cell survival rate after sorting was (95.70±2.02)%, and the cell morphology was good.Conclusion The flow cytometer can quickly and efficiently isolate NK cells from mouse lungs, and the sorted cells maintain good viability, which can provide a guarantee for further research on NK cells.

参考文献/References:

[1]Calabrese DR,Aminian E,Mallavia B,et al.Natural killer cells activated through NKG2D mediate lung ischemia-reperfusion injury[J].J Clin Invest,2020,12(8):137047. [2]Parihar R,Rivas C,Huynh M,et al.NK Cells Expressing a Chimeric Activating Receptor Eliminate MDSCs and Rescue Impaired CAR-T Cell Activity against Solid Tumors[J].Cancer Immunol Res,2019,7(3):363-375. [3]Stokic-Trtica V,Diefenbach A,Klose CSN.NK Cell Development in Times of Innate Lymphoid Cell Diversity[J].Front Immunol,2020(11):813. [4]Tu Y,Pan M,Song S,et al.CD3+ CD56+ natural killer T cell infiltration is increased in cervical cancer and negatively correlated with tumour progression[J].Biotechnology & Biotechnological Equipment,2019,33(1):1380-1391. [5]Becker PS,Suck G,Nowakowska P,et al.Selection and expansion of natural killer cells for NK cell-based immunotherapy[J].Cancer Immunol Immunother,2016,65(4):477-484. [6]Kloβ S,Oberschmidt O,Morgan M,et al.Optimization of Human NK Cell Manufacturing: Fully Automated Separation, Improved Ex Vivo Expansion Using IL-21 with Autologous Feeder Cells, and Generation of Anti-CD123-CAR-Expressing Effector Cells[J].Hum Gene Ther,2017,28(10):897-913. [7]Guillerey C.NK Cells in the Tumor Microenvironment[J].Adv Exp Med Biol,2020(1273):69-90. [8]Wang G,Yu G,Wang D,et al.Comparison of the purity and vitality of natural killer cells with different isolation kits[J].Exp Ther Med,2017,13(5):1875-1883. [9]Alvarez M,Ochoa MC,Minute L,et al.Rapid isolation and enrichment of mouse NK cells for experimental purposes[J].Methods Enzymol,2020(631):257-275. [10]卢铨,陈浩,沙卫红,等.从健康人外周血单个核细胞中分离出高纯度的CD3-CD56+CD16+NK细胞对肝癌细胞的杀伤作用分析[J].胃肠病学和肝病学杂志,2019,28(9):978-982. [11]Holicek P,Truxova I,Kasikova L,et al.Assessment of NK cell-mediated cytotoxicity by flow cytometry after rapid, high-yield isolation from peripheral blood[J].Methods Enzymol,2020(631):277-287. [12]程明,孙汭,田志刚,等.人类肝脏NK 细胞与肝脏疾病[J].中国免疫学杂志,2020,36(11)1281-1290. [13]Montaldo E,Del Zotto G,Della Chiesa M,et al. Human NK cell receptors/markers: a tool to analyze NK cell development, subsets and function[J].Cytometry A,2013,83(8):702-713. [14]杨梅.CD11b与CD27定义的人类NK细胞亚群在HIV疾病进展中的变化及相关机制的研究[D].中国医科大学,2017. [15]Yudhistira T,Mulay SV,Kim Y,et al.Imaging of Hypochlorous Acid by Fluorescence and Applications in Biological Systems[J].Chem Asian J,2019,14(18):3048-3084. [16]Dunker S.Hidden Secrets Behind Dots: Improved Phytoplankton Taxonomic Resolution Using High-Throughput Imaging Flow Cytometry[J].Cytometry A,2019,95(8):854-868.

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更新日期/Last Update: 1900-01-01