Evidence for the presence of viable endothelial cells in cultures derived from dissociated rat brain

Abstract

The morphology and histochemistry of dissociated newborn rat brain was studied in tissue culture. Direct microscopy of developing cells, electron microscopy and the alkaline phosphatase activity were used to identify the capillary endothelial cells.

Rat Cerebellar Cells in Tissue Culture

“…The most widely used methods of determining the identity of these cells have been histochemi cal detection of alkaline phosphatase and yglutamyl transpeptidase activities [10,15,22,28]. We have confirmed that endothelial cells in cryostat sections do indeed appear to con tain much higher levels of alkaline phospha tase activity than the neural cell types, al though electron microscopic studies have shown that the enzyme is not found solely in endothelial cells [29].…”

“…Several authors have recently reported the growth of brain endothelial cells in dis persed cell culture [9,10,15,22,28]. The most widely used methods of determining the identity of these cells have been histochemi cal detection of alkaline phosphatase and yglutamyl transpeptidase activities [10,15,22,28].…”

“…It has been proposed that alkaline phos phatase and y-glutamyl transpeptidase activ ity can be regarded as characteristic markers for endothelial cells of brain capillaries [22,28]. With this in mind we investigated the histochemical distribution of alkaline phos phatase in both cryostat sections and cell cul tures of the cerebellum.…”

“…While sev eral laboratories have recently reported the growth of endothelial cells in culture [9. 15, 22, 28] their unequivocal identification re mains a problem since enzymes characteristic of these cells are not always expressed in vi tro [10,22,28]. It has been shown by Cossio et al [7,8] using the indirect immunofluorescent technique that sera from patients with Chagas' disease (a South American trypano-somiasis caused by the protozoan Trypano soma cruzi) contain immunoglobulins which bind both to the interstitium and blood ves sels of heart and skeletal muscle, to heart en docardium, and to vascular structures of liver, kidney, stomach and placenta.…”

Recent advances in understanding brain capillary function

“…During the past few years, several laboratories have developed methods for culturing endothelial cells from brain microvessels 11 1, 24, 63,68,76). In this issue of the Annals, Bowman and colleagues 112) describe some of the features of cultured endothelial cells prepared from bovine brain.…”