Construction of Blood-brain Barrier in vitro Model Based on Microfluidic Chip

doi:10.13523/j.cb.20171201

China Biotechnology

2017, Vol. 37

Issue (12): 1-7 DOI: 10.13523/j.cb.20171201

Orginal Article

Construction of Blood-brain Barrier in vitro Model Based on Microfluidic Chip

Li-li JIANG,Jun-song ZHENG,Yan LI,Jun DENG,Li-chao FANG,Hui HUANG(

)

Department of Clinical Laboratory Medicine,The First Affiliated Hospital,Third Military Medical University ,Chongqing 400038,China

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Abstract Objective:

To create a blood-brain barrier in vitro model by microfluidic chip technology, which were easy to control and close to microenvironment.

Methods:

The microfluidic chips were composed of dual pool structure, separated by the polycarbonate membrane, and there were two sets of flow control system, mouse brain vascular endothelial cells and astrocytes were isolated and purified, the cells were identified by immunofluorescence. Then cells were added to the two pools according to the order, and infused at 1μl/min flow rate. The blood-brain barrier(BBB) model was constructed, and then identified and evaluated.

Results:

Two kinds of cells were obtained by isolation and purification, and the cells were identified by immunofluorescence and the purity was above 95%. After 3 days of co-culture, tight junctions began to develop, and the peak is on the fifth day, the ultrastructural observation showed that the mouse brain vascular endothelial cells formed tight junctions, the penetration experiments of fluorescein sodium and the measurement of TEER showed that barrier formation was good.

Conclusion:

The blood-brain barrier model was constructed based on the microfluidic chip, It may become a new platform for drug screening, neural system basis and other studies.

Key words： Microfluidic chip Blood-brain barrier model Microenvironment

Received: 05 July 2017 Published: 16 December 2017

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	Li-li JIANG
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	Hui HUANG

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Li-li JIANG,Jun-song ZHENG,Yan LI,Jun DENG,Li-chao FANG,Hui HUANG. Construction of Blood-brain Barrier in vitro Model Based on Microfluidic Chip. China Biotechnology, 2017, 37(12): 1-7.

URL:

https://manu60.magtech.com.cn/biotech/10.13523/j.cb.20171201 OR https://manu60.magtech.com.cn/biotech/Y2017/V37/I12/1

Fig.1 Design and fabrication of BBB model for microfluidic chip
(a) Chip design upper layer (b) Chip design lower layer (c) The diagram of chip mounting (d) The diagram of cells with chip (e)Silicon mold (f) Microfluidic device system

Fig.2 Cell preparation of the BBB model based on the microfluidic chip
(a) Cerebral microvascular endothelial cells were cultured for 7 days (×100) (b) vWF staining ,DAPI staining (×200) (c) Astrocytes were purified for fourth generations (×100) (d) GFAP staining ,DAPI staining (×200)

Fig.3 Transmission electron microscope of BBB model based on microfluidic chip
(a) Two layers of cells were co-culture and separated by PC membranes (b)The tight junction protein of the cerebral vascular endothelial cells

Fig.4 The penetration experiments of fluorescein sodium with BBB model

Fig.5 The TEER measurement of BBB model


[1]	刘超, 李明昌, 陈谦学 , 等. 血脑屏障结构与功能及其在缺血性脑血管病中的研究进展. 国际神经病学神经外科学杂志, 2016,43(6):564-568 .

[1]	Liu C, Li M C, Chen Q X , et al. The structure and function of blood-brain barrier and its progress in ischemic cerebrovascular disease. Journal of International Neurology and Neurosurgery, 2016,43(6):564-568.

[2]	林晓梅, 张铭 . PDMS微流控芯片加工技术研究. 长春工业大学学报, 2013,34(2):172-179.

[2]	Lin X M, Zhang M . Experimental study on fabrication of PDMS. Journal of Changchun University of Technology, 2013,34(2):172-179.

[3]	伊波立 . Tat 肽段介导的超顺磁性纳米铁颗粒通过体外血脑屏障模型的研究. 武汉:华中科技大学, 2010. 11-13. doi: 10.7666/d.d140125

[3]	Yi B L . The Research of Tat peptide-assisting USPIO crossing the blood brain barrier model. Wuhan:Huazhong University of Science and Technology, 2010. 11-13. doi: 10.7666/d.d140125

[4]	李珺, 彭亮, 黄胜和 , 等. 体外血脑屏障模型的建立和发展. 广东医学, 2009,30(4):647-648. doi: 10.3969/j.issn.1001-9448.2009.04.068

[4]	Li J, Peng L, Huang S H , et al. Establishment and development of blood-brain barrier model in vitro. Guangdong Medical Journal, 2009,30(4):647-648 doi: 10.3969/j.issn.1001-9448.2009.04.068

[5]	Helms H C, Abbott N J, Burek M , et al. In vitro models of the blood-brain barrier: An overview of commonly used brain endothelial cell culture models and guidelines for their use. J Cereb Blood Flow Metab, 2016,36(5):862-890. doi: 10.1177/0271678X16630991

[6]	Chen L, Zheng X L, Ning H U , et al. Research progress on microfluidic chip of cell separation based on Dielectrophoresis. Chinese Journal of Analytical Chemistry, 2015,43(2):300-309. doi: 10.1016/S1872-2040(15)60808-8

[7]	Decrop D, Brans T, Gijsenbergh P , et al. Optical manipulation of single magnetic beads in a microwell array on a digital microfluidic chip. Analytical Chemistry, 2016,88(17):8596-8612. doi: 10.1021/acs.analchem.6b01734 pmid: 27448015

[8]	Jia M L, Meng Z, Yeong W Y . Characterization and evaluation of 3D printed microfluidic chip for cell processing. Microfluidics and Nanofluidics, 2016,20(1):1-15. doi: 10.1007/s10404-015-1676-z

[9]	Helm M, Meer A, Eijkel J , et al. Microfluidic organ-on-chip technology for blood-brain barrier research. Tissue Barriers, 2016,4(1):1-13. doi: 10.1080/21688370.2016.1142493 pmid: 27141422

[10]	Jia M L, Meng Z, Yeong W Y . Characterization and evaluation of 3D printed microfluidic chip for cell processing. Microfluidics and Nanofluidics, 2016,20(1):1-15. doi: 10.1007/s10404-015-1676-z

[11]	Caplin J D, Granados N G, James M R , et al. Microfluidic organ-on-a-chip technology for advancement of drug development and toxicology. Advanced Healthcare Materials, 2015,4(10):1426-1431. doi: 10.1002/adhm.201500040 pmid: 25820344

[12]	Fu Y, Zhou H, Jia C , et al. A microfluidic chip based on surfactant-doped polydimethylsiloxane (PDMS) in a sandwich configuration for low-cost and robust digital PCR. Sensors & Actuators B Chemical, 2017,245:414-422. doi: 10.1016/j.snb.2017.01.161

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