Permeability properties of a three-cell type in vitro model of blood-brain barrier

Schiera, Gabriella; Sala, Sandra; Gallo, Alessia; Raffa, Maria Pia; Pitarresi, Giovanna; Savettieri, Giovanni; Liegro, Italia Di

doi:10.1111/j.1582-4934.2005.tb00362.x

Cited by 62 publications

(58 citation statements)

References 20 publications

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“…The presence of both astrocytes and neurons decreased the paracellular permeability of RBE4 immortalized rat brain endothelial cells (Schiera et al 2005). In a flow-based in vitro BBB model, the differentiation of serotonergic neurons was promoted by the endothelial-glial co-culture (Stanness et al 1999).…”

Section: Resultsmentioning

confidence: 99%

Pericytes from Brain Microvessels Strengthen the Barrier Integrity in Primary Cultures of Rat Brain Endothelial Cells

et al. 2007

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(1) The blood-brain barrier (BBB) characteristics of cerebral endothelial cells are induced by organ-specific local signals. Brain endothelial cells lose their phenotype in cultures without cross-talk with neighboring cells. (2) In contrast to astrocytes, pericytes, another neighboring cell of endothelial cells in brain capillaries, are rarely used in BBB co-culture systems. (3) Seven different types of BBB models, mono-culture, double and triple co-cultures, were constructed from primary rat brain endothelial cells, astrocytes and pericytes on culture inserts. The barrier integrity of the models were compared by measurement of transendothelial electrical resistance and permeability for the small molecular weight marker fluorescein. (4) We could confirm that brain endothelial monolayers in mono-culture do not form tight barrier. Pericytes induced higher electrical resistance and lower permeability for fluorescein than type I astrocytes in co-culture conditions. In triple co-culture models the tightest barrier was observed when endothelial cells and pericytes were positioned on the two sides of the S. Nakagawa Á M. A. Deli Á S. Nakao Á R. Nakaoke Á M. Niwa Department of Pharmacology, Nagasaki University Graduate School of Biomedical Sciences, 1-12-4 Sakamoto, Nagasaki 852-8523, Japan 687-694 DOI 10.1007/s10571-007-9195-4 123 porous filter membrane of the inserts and astrocytes at the bottom of the culture dish. (5) For the first time a rat primary culture based syngeneic triple co-culture BBB model has been constructed using brain pericytes beside brain endothelial cells and astrocytes. This model, mimicking closely the anatomical position of the cells at the BBB in vivo, was superior to the other BBB models tested. (6) The influence of pericytes on the BBB properties of brain endothelial cells may be as important as that of astrocytes and could be exploited in the construction of better BBB models.

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Section: Resultsmentioning

confidence: 99%

Pericytes from Brain Microvessels Strengthen the Barrier Integrity in Primary Cultures of Rat Brain Endothelial Cells

et al. 2007

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“…Although useful these methods have not been widely adopted, likely due to their low yields. As such, most studies characterizing neural/endothelial cell interactions continue to rely on immortalized cell lines that may exhibit altered properties due to their in vitro derivation and passage [10,16].…”

Section: Discussionmentioning

confidence: 99%

“…[5][6][7][8][9][10][11][12][13][14][15][16]. This method utilizes a mild trypsinization of embryonic brain that permits the sieve separation of neural cells and the subsequent liberation of EC from cerebral vessels with a gentle collagenase/dispase treatment.…”

Section: Characterization Of Embryonic Vascular Endothelial Cells (Ec)mentioning

confidence: 99%

Interactions of primary neuroepithelial progenitor and brain endothelial cells: distinct effect on neural progenitor maintenance and differentiation by soluble factors and direct contact

et al. 2007

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Neurovascular interactions are crucial for the normal development of the central nervous system. To study such interactions in primary cultures, we developed a procedure to simultaneously isolate neural progenitor and endothelial cell fractions from embryonic mouse brains. Depending on the culture conditions endothelial cells were found to favor maintenance of the neuroprogenitor phenotype through the production of soluble factors, or to promote neuronal differentiation of neural progenitors through direct contact. These apparently opposing effects could reflect differential cellular interactions needed for the proper development of the brain.

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“…In order to set the BBB in vitro model, we used the transwell system described elsewhere (23)(24)(25). Briefly, neurons were cultured on laminin at the bottom of the wells of the companion plates, and fed with MM for 5 days.…”

Section: Methodsmentioning

confidence: 99%

Neuronal and BBB damage induced by sera from patients with secondary progressive multiple sclerosis

Proia

Schiera²,

Salemi³

et al. 2009

Int J Mol Med

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Abstract. An important component of the pathogenic process of multiple sclerosis (MS) is the blood-brain barrier (BBB) damage. We recently set an in vitro model of BBB, based on a three-cell-type co-culture system, in which rat neurons and astrocytes synergistically induce brain capillary endothelial cells to form a monolayer with permeability properties resembling those of the physiological BBB. Herein we report that the serum from patients with secondary progressive multiple sclerosis (SPMS) has a damaging effect on isolated neurons. This finding suggests that neuronal damaging in MS could be a primary event and not only secondary to myelin damage, as generally assumed. SPMS serum affects the permeability of the BBB model, as indicated by the decrease of the transendothelial electrical resistance (TEER). Moreover, as shown by both immunofluorescence and Western blot analyses, BBB breaking is accompanied by a decrease of the synthesis as well as the peripheral localization of occludin, a structural protein of the tight junctions that are responsible for BBB properties.

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Permeability properties of a three-cell type in vitro model of blood-brain barrier

Cited by 62 publications

References 20 publications

Pericytes from Brain Microvessels Strengthen the Barrier Integrity in Primary Cultures of Rat Brain Endothelial Cells

Pericytes from Brain Microvessels Strengthen the Barrier Integrity in Primary Cultures of Rat Brain Endothelial Cells

Interactions of primary neuroepithelial progenitor and brain endothelial cells: distinct effect on neural progenitor maintenance and differentiation by soluble factors and direct contact

Neuronal and BBB damage induced by sera from patients with secondary progressive multiple sclerosis

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