|本期目录/Table of Contents|

[1]陈 丹,王小东,童静植,等. 三种分离人外周血单核细胞方法的比较[J].天津医科大学学报,2014,20(06):483-485.
 CHEN Dan,WANG Xiao-dong,TONG Jing-zhi,et al.Comparison of three methods of separating human peripheral blood mononuclear cells [J].Journal of Tianjin Medical University,2014,20(06):483-485.
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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:
20
期数:
2014年06期
页码:
483-485
栏目:
技术与方法
出版日期:
2014-11-20

文章信息/Info

Title:

Comparison of three methods of separating human peripheral blood mononuclear cells

文章编号:
1006-8147(2014)06-0485-03
作者:
陈 丹1 王小东2 童静植2 丁 文2 孙 科2 何景华1 强兆艳1 李 光3 康 毅1
 (天津医科大学1.基础医学院药理学教研室;2. 2009级临床医学七年制;3.基础医学院生物学教研室,天津300070)
Author(s):

 CHEN Dan1WANG Xiao-dong2TONG Jing-zhi2DING Wen2SUN Ke2HE Jing-hua1 QIANG Zhao-yan1LI Guang3KANG Yi1

(1.Department of Pharmacology; 2.Clinical Seven-years Students, Grade 2009; 3.Department of Biology, Tianjin Medical University, Tianjin 300070, China)
关键词:
 单核细胞贴壁法免疫磁珠流式细胞术
Keywords:
monocyte adherent separation method immune magnetic beads separation flow cytometry separation
分类号:
R446
DOI:
-
文献标志码:
A
摘要:
 目的:用3种方法从人外周血分离单核细胞,比较细胞纯度、得率、实验成本及所需时间,选择最适宜的分离方法。方法:用密度梯度离心法获得外周血的单个核细胞,分别用贴壁法、免疫磁珠法和流式细胞术法进一步分离单核细胞。经CD14抗体标记,用流式细胞术检测细胞纯度,计算细胞得率、实验成本及所需时间。结果:用贴壁法、免疫磁珠法和流式细胞术法分离单核细胞,细胞纯度分别为18.8%、80%和73.4%;细胞得率为5%、11%和7.5%;每获得2×107个单核细胞,实验成本约500元、1 500元和2 100元,实验耗时约6 h、6 h和10 h。结论:免疫磁珠法是最适宜的分离人外周血单核细胞的方法,具有细胞纯度与得率较高,实验成本较低,耗时较短的优点。
Abstract:
Objective: To separate monocyte from human peripheral blood based on cell purity, cell yield rate, experiment cost and the time required. Methods: Peripheral blood mononuclear cells were obtained via density gradient centrifugation method. Then the adherent method, immune magnetic beads method and flow cytometry method were used to separate monocyte from peripheral blood mononuclear cells. Cell purity was detected by flow cytometry after being marked with CD14 monoclonal antibody. Cell yield rate, experiment cost and the time required were calculated. Results: The cells purity was 18.8% by adherent method while cells purity was 80% and 73.4% respectively indicated by immune magnetic beads and flow cytometry methods. The cell yield rates were 5%, 11% and 7.5%, respectively. The experiment costs were about 500 yuan, 1500 yuan and 2100 yuan every 2×107 mononuclear cells while the time required for experiment were about 6 h, 6 h and 10 h, respectively. Conclusion: Immune magnetic beads method is the most appropriate method to separate monocyte, with the advantages of higher cell purity and yield rate, lower experiment cost and shorter time as compared with other methods.

参考文献/References:

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[2] 马斌,曲萍,张秀敏. 连续贴壁法分离培养人外周血来源树突状细胞及其超微结构的观察[J]. 现代肿瘤医学. 2007,15(3):0301

[3] 吴鹏,刘映峰,梁东辉,等. 提高人外周血单核细胞分离率的方法讨论[J].实用医学杂志,2008,24(5):707

[4] 陈媛,巩伟丽,湛小燕等. 应用流式细胞分选技术分离人外周血原代单核细胞[J]. 科学技术与工程,2012,12(24):5985

[5] 韩亚萍,刘源,章莉莉等. 外周血单核细胞的分离方法探讨[J]. 中华现代临床医学杂志,2003,1(9):780

[6] 刘剑雯,曾志勇,陈君敏. 人外周血耐受性树突状细胞的诱导培养及其DC-STAMP的表达研究[J]. 中国免疫学杂志,2013,(5):474

[7] 徐勇,霍梅.免疫磁珠分离及流式细胞仪分选纯化外周血 CD34+/CD90+干细胞[J]. 临床检验杂志,2004,22(4):246

[8] 郭陈智,史桂英. 流式分选仪FACSAria II分选GFP阳性乳腺癌MCF-7细胞株的条件优化研究[J]. 上海交通大学学报,2013,33(1):127

[9] 张玮玮.流式细胞术的发展、原理及临床应用[J].当代医学,2008,(2):25

[10] 张锦英,束永前.流式细胞术的工作原理及临床应用[J].中国生化药物杂志, 2003,24(6):308

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备注/Memo

备注/Memo:

基金项目 国家高新技术研究发展计划(863计划)第八课题组子课题(NO.2012AA021003)

作者简介 陈丹(1980-),女,副教授,博士,研究方向:中药药理学;通信作者:李光,E-mail:lig@tijmu.edu.cn,康毅, E-mail:13902075982@163.com



更新日期/Last Update: 2014-12-03