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P.1 PBECs or porcine brain microvessels were washed with PBS, fixed with 3% paraformaldehyde for 45 min and then permeabilised in 0.1% Triton X-100.
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Microvessels were washed of blood cells by filtering through a 40 μm cell strainer (Becton Dickinson Labware, Franklin Lakes, IN, USA) and eluting with PBS.
Digested fragments of microvessels were washed and then seeded onto 60-mm-diameter dishes coated with Matrigel (Matrigel growth factor reduced, Becton Dickinson; dilution 1/50 in DMEM).
Brain microvessels were obtained as described previously [18].
To explain Aβ enhanced clearance across the BBB, brain microvessels were isolated for analysis.
13 After removing the leptomeninges from the brain, microvessels were isolated out of the grey matter.
Brain microvessels were isolated as described previously (Seelbach et al. 2007).
All steps in the isolation of brain microvessels were carried out at 4°C.
Brain microvessels were isolated according to the method of Yamakawa et al. [ 17].
In the present study, isolated brain microvessels were used to examine the expression of proteins that mediate angiogenesis and accurately corresponded to the observed alterations in the brain microvessels; significantly lower VEGF expression was observed in the aging rats.
Brain microvessels were isolated from naïve mice and mice infected 7 days earlier with PbA, PbNK65 or Py17X.
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