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whether passive or active transport dispersion process is found similar in vivo [2]. Because
of these characteristics, this model system possesses many advantages over conventional
in vivo studies, for example, rapid assessment of invasion of pathogenic microorganism
and proteins, genes, cell signaling pathways which have business with bacterium invasion.
This in vitro BBB model system would have a wide application for microbial meningitis
and other studies about how pathogenic microbe destroy or pass through the BBB without
expensive animals studies.
There were many discussed methods for isolations of BMEC were reported in the
past three decades, and the most common means of isolation include homogenate, filtra-
tion, centrifuge and digestion. Besides, there were labs use tissue culture and white
matter of brain for culture[3,6,9~15]. However, spending long time, big workload de-
manding instruments, survival difficulties of BMEC and growth of mixed fibroblasts are
still common problems. How to cultivate endothelial cells with high purity and charac-
teristics of the BBB, is always a difficulty for researchers no matter interior or abroad.
A method about separation and cultivate successfully for the brain microvascular en-
dothelial cells with high purity and growth activity would be reported by this article.
Fig. 1. The BBB structure: Brain microvascular endothelial cell is the main component of the
BBB. The BBB in vivo is characterized by tight intercellular junction, few pinocytic vesicles,
and lacking fenestra. (Source: Sheng-He Huang and Ambrose Y. Jong. Cellular mechanisms of
microbial proteins contributing to invasion of the blood-brain barrier).
2 Materials and Methods
2.1 Animals
Eight rats about two- to three-week-old.
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