[1]宋晓彬,范春娥,王 烨,等.小鼠脑微血管内皮细胞的分离培养与鉴定[J].南京师范大学学报(自然科学版),2017,40(03):128.[doi:10.3969/j.issn.1001-4616.2017.03.021]
　Song Xiaobin,Fan Chune,Wang Ye,et al.Primary Culture and Identification of Mouse BrainMicrovascular Endothelial Cell[J].Journal of Nanjing Normal University(Natural Science Edition),2017,40(03):128.[doi:10.3969/j.issn.1001-4616.2017.03.021]
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小鼠脑微血管内皮细胞的分离培养与鉴定()
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《南京师范大学学报》（自然科学版）[ISSN:1001-4616/CN:32-1239/N]

卷:
第40卷
期数:
2017年03期
页码:
128
栏目:
·生命科学·
出版日期:
2017-09-30

文章信息/Info

Title:
Primary Culture and Identification of Mouse BrainMicrovascular Endothelial Cell
文章编号:
1001-4616(2017)03-0138-06
作者:
宋晓彬1; 范春娥1; 王 烨2; 张艺腾1; 崔丛丛1; 曾卫卫1; 郑丽蓉1; 蔡晓洁1; 陈华群1
(1.南京师范大学生命科学学院,江苏省分子医学生物技术重点实验室,江苏 南京 210023)(2.南京沐赛生物技术有限公司,江苏 南京 210061)
Author(s):
Song Xiaobin1; Fan Chun’e1; Wang Ye2; Zhang Yiteng1; Cui Congcong1; Zeng Weiwei1; Zheng Lirong1; Cai Xiaojie1; Chen Huaqun1
(1.School of Life Sciences,Nanjing Normal University,Jiangsu Key Laboratory for Molecular and Medical Biotechnology,Nanjing 2l0023,China)(2.Nanjing Mucyte Biotechnical Com.,Nanjing 210061,China)
关键词:
脑微血管内皮细胞; 原代培养; 密度梯度离心; 酶消化
Keywords:
mouse brain microvescular endothelial cell; primary culture; density gradient centrifugation; enzyme digestion
分类号:
Q28
DOI:
10.3969/j.issn.1001-4616.2017.03.021
文献标志码:
A
摘要:
脑微血管内皮细胞(Brain microvascular endothelial cell,BMVEC)是构成血脑屏障(Brain-blood barrier,BBB)的基本组分,多种神经疾病都与BBB的失功能相关,分离培养原代BMVEC是体外研究BBB功能及其调节的重要手段. 本文通过简单有效的葡聚糖密度梯度离心和酶消化方法,从小鼠脑组织分离培养出BMVEC,并用免疫荧光染色进行了鉴定. 结果表明,内皮细胞表面特征性的标记分子CD31阳性细胞达到95%以上,同时通过VE-Cadherin及ZO-1免疫荧光染色鉴定证实,其为脑微血管内皮细胞. 本研究为体外利用BMVEC进行相关研究提供了良好基础.
Abstract:
Brain microvascular endothelial cell(BMVEC)is an essential component of the brain blood barrier(BBB). It has been known that the dysfunction of BBB plays critical roles in the pathological process of a variety of neurological disorders. The primary cultured BMVECs were widely used in the study of the functions and the regulation mechanisms of the BBB. Here,it is reported that a simple and effective method of isolating and growing of BMVEC established upon density gradient centrifugation and enzyme digestion. The purity of the cultured cells was higher than 95% determined by CD31 immunofluorescence staining. The features of the cells were further confirmed by positive staining of VE-cadherin and ZO-1,the characteristics of BMVECs. Our method may provide a good approach for the study of BBB in vitro.

参考文献/References:

[1] BALLABH P,BRAUN A,NEDERGAARD M. The blood-brain barrier:an overview:structure,regulation,and clinical implications[J]. Neurobiology of disease,2004,16(1):1-13.
[2]NEUWELT E A,BAUER B,FAHLKE C,et al. Engaging neuroscience to advance translational research in brain barrier biology[J]. Nature reviews neuroscience,2011,12(3):169-182.
[3]ZOLEZZI J M,INESTROSA N C. Peroxisome proliferator-activated receptors and Alzheimer’s disease:hitting the blood-brain barrier[J]. Molecular neurobiology,2013,48(3):438-451.
[4]SHI Y,ZHANG L,PU H,et al. Rapid endothelial cytoskeletal reorganization enables early blood-brain barrier disruption and long-term ischaemic reperfusion brain injury[J]. Nature communications,2016,7(10):523.
[5]MINAMINO T,MIYAUCHI H,YOSHIDA T,et al. Endothelial cell senescence in human atherosclerosis role of telomere in endothelial dysfunction[J]. Circulation,2003,105(13):1 541-1 544.
[6]ZLOKOVIC B V. The blood-brain barrier in health and chronic neurodegenerative disorders[J]. Neuron,2008,57(2):178-201.
[7]OHTSUKI S,TERASAKI T. Contribution of carrier-mediated transport systems to the blood-brain barrier as a supporting and protecting interface for the brain; importance for cns drug discovery and development[J]. Pharmaceutical research,2007,24(9):1745-1758.
[8]OBERMEIER B,DANEMAN R,RANSOHOFF R M. Development,maintenance and disruption of the blood-brain barrier[J]. Nat Med,2013,19(12):1584-1596.
[9]PERSIDSKY Y,RAMIREZ S H,HAORAH J,et al. Blood-brain barrier:structural components and function under physiologic and pathologic conditions[J]. Journal of neuroimmune pharmacology,2006,1(3):223.
[10]BETZ A L,GOLDSTEIN G W. Primary culture of capillary endothelium from rat brain[J]. In vitro cellular and developmental biology-plant,1981,17(4):353-362.
[11]ABBOTT N J,HUGHES C C,REVEST P A,et al. Development and characterisation of a rat brain capillary endothelial culture:towards an in vitro blood-brain barrier[J]. Journal of cell science,1992,103(1):23-37.
[12]NAKAGAWA S,DELI M A,NAKAO S,et al. Pericytes from brain microvessels strengthen the barrier integrity in primary cultures of rat brain endothelial cells[J]. Cell Mol Neurobiol,2007,27(6):687-694.
[13]许熊飞,李润平,李泉,等. 大鼠脑微血管内皮细胞的分离与原代培养[J]. 中国细胞生物学学报,2005,27(1):84-88.
[14]NAVONE S E,MARFIA G,INVERNICI G,et al. Isolation and expansion of human and mouse brain microvascular endothelial cells[J]. Nature protocols,2013,8(9):1680.
[15]WU Z,HOFMAN F M,ZLOKOVIC B V. A simple method for isolation and characterization of mouse brain microvascular endothelial cells[J]. Journal of neuroscience methods,2003,130(1):53-63.
[16]林娜,王安平. 小鼠脑血管内皮细胞的分离和培养[J]. 南昌大学学报(医学版),2003,43(6):52-53.
[17]朱玉珍,武文,田野苹,等. 小鼠脑微血管内皮细胞的原代培养与纯化[J]. 解剖学杂志,2007,30(5):530-533.
[18]张畅斌,陆茵,李沧海,等. 小鼠脑微血管内皮细胞的培养及其生物学行为初探[J]. 南京中医药大学学报(自然科学版),2012,28(2):139-142.
[19]徐剑文,王玮,康仲涵,等. 小鼠脑微血管内皮细胞的体外培养[J]. 福建医科大学学报(自然版),2000,34(3):215-217.

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备注/Memo

备注/Memo:
收稿日期:2017-04-18.
基金项目:国家自然科学基金(31371356、30971540)、江苏省教育厅重大项目(13KJA180004).
通讯联系人:陈华群,教授,研究方向:细胞生物学. E-mail:chenhuaqun@njnu.edu.cn
更新日期/Last Update: 2017-09-30