Cerebral microvessels and derived cells in tissue culture: Isolation and preliminary characterization

Abstract

Microvessels isolated from mouse forebrain were used as the source material for the derivation of cerebral vascular endothelium and smooth-muscle cells in culture. The microvessels were isolated by a mechanical dispersion and filtration technique, and were maintained in vitro as organoid cultures. A microvessel classification system was developed and proved to be useful as a tool in monitoring culture progress and in predicting the type(s) of microvessel(s) that would give rise to migrating and/or proliferating cells. The isolated cerebral microvessels were heterogeneous in diameter, size of individual vascular isolate, and proliferative potential. The isolated microvessels ranged in diameter from 4 micron to 25 micron and in size from a single microvascular segment to a large multibranched plexus with mural cells. The initial viability, determined by erythrosin B exclusion, was approximately 50% on a per cell basis. All microvessel classes had proliferative potential although the rate and extent of proliferation were both microvessel class- and density-dependent. The smaller microvessels gave rise to endothelial cells, whereas the large microvessels gave rise to endothelial and smooth-muscle cells. The viability and progress of a microvessel toward derived cell proliferation seemed to be directly proportional to the number of mural cells present.

Arachidonic acid and prostaglandin endoperoxide metabolism in isolated rabbit and coronary microvessels and isolated and cultivated coronary microvessel endothelial cells.

“…Removal of the medium and unattached cells 30 min after inoculation helped to select for endothelial cells. Although previous studies with capillary endothelium have reported the requirement for gelatin, collagen, or other substratum materials (19,27), in our hands the RCME cells attached as well to untreated culture plates. The primary cultures required high serum concentrations (15-20%) to support growth.…”

“…In the past 3 to 4 yr, rapid progress has been made in the isolation and culture of microvascular endothelial cells (17)(18)(19)(20)(21)(22)(23)(24)(25). These studies have demonstrated that microvascular endothelial cells exhibit several fundamental differences potentially useful for the understanding and treatment of diseases with a microvascular component.…”

Monoamine Oxidase Activity in the Cerebral Vasculature: Comparison between Fresh Microvessels from Different Structures and Cell Cultures Derived from Microvessels

“…The procedure used for the isolation of the microves sels was derived from the method of Debault et al (1979). The cortex, cleared of the meninges, was cut into small pieces and washed with a phosphate-buffered salt solution (PBS) of pH 7.0 containing 2 giL glucose and 0.…”

Differential adhesion of tumor cells to capillary endothelial cells in vitro.

“…[6][7][8] Cell Cultures. The mouse brain endothelial cell line MBE-1 (called MBE), isolated in our laboratory by J. Joseph (4,5,13), was obtained from cerebral microvessels of strain A.TL mice according to published protocols of DeBault et al (14). After initial culture in modified Lewis medium (14), the line was maintained in Dulbecco's modification of Eagle's minimum essential medium (DME medium) supplemented with 15-20% fetal bovine serum and 20% S-180 tumor-conditioned medium as described by Folkman et al (15).…”

“…FB-356 fibroblasts exposed to 5000 rad of y-irradiation served as feeder layers for teratoma cell lines. M-5076 cells were grown in DeBault's medium (14) but could be maintained in primary culture only.…”