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The Establishment of in vitro Model of Primary Rat
Brain Microvascular Endothelial Cells
Jun Li 1 , Liang Peng 1 , Xiao-Lu Liu 1 , Hong Cao 1, * ,
Sheng-He Huang 1,2, * , and Chun-Hua Wu 2
School of Public Health and Tropical Medicine, Southern Medical University,
Guangzhou, Guangdong 510515, China
2 Children's Hospital Los Angeles, the University of Southern California,
Los Angeles, USA
{Jun Li,Liang Peng,Xiao-Lu Liu,Hong Cao},
{Sheng-He Huang,Chun-Hua Wu}
Abstract. In order to establish an in vitro brain microvascular endothelial cells
(BMEC) model of blood-brain barrier (BBB), we isolated and cultivated pri-
mary microvascular endothelial cells from grey matters of rat brain. The grey
matter was squashed into homogenate, centrifugalized with dextran, and di-
gested. Then the microvessels were seeded onto collagen-coated 25cm 2 tissue
culture flasks. Then the cell was examined with microscope. Microscopically
the isolated cells in the fusiform, when cultured in 24 hour, formed the cobble-
stone shape. These cells reached confluence in the form of uniform monolayer
after 7 days in culture. Therefore, the primary BMECs of rat brain were suc-
cessfully isolated and cultivated by using the above method, and we could use
this model to study the relationship between BMEC and pathogenic microbes.
Keywords: Rat, Primary culture, BMEC.
1 Introduction
The first successful isolation of brain microvessels from animal brain was 30 years
agoand then many scientists studied several new ways to develop in vitro models
system of the blood-brain barrier (BBB), in order to establish a more stable and simi-
lar to the blood-brain barrier in vivo . And from then on, these BBB model systems in
vitro have contributed considerably to the understanding of the physiology, pharma-
cology, pathophysiology and centre nervous system infection of the BBB.
This Brain microvascular endothelial cell (BMEC) is the main component of blood-
brain barrier (BBB). The BMEC monolayer in vitro model system are morphologically
similar to endothelial cells of the BBB in vivo, characterized by tight intercellular junction,
few pinocytic vesicles, and lacking fenestra. Markers for cells of endothelial origin, for
example, angiotensin-converting enzyme, and factor VIII antigen, have been also demon-
strated in this model[1]. The transfer function studies on the model in vitro showed that
* Corresponding authors.
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