A Novel Brain Neurovascular Unit Model with Neurons, Astrocytes and Microvascular Endothelial Cells of Rat

Abstract

A novel triple cell neurovascular unit (NVU) model co-culturing with neurons, brain microvascular endothelial cells (BMECs) and astrocytes was established in this study for investigating the cerebral diseases and screening the candidates of therapeutic drug. We have first performed the cell identification and morphological characterization, analyzed the specific protein expression and determined the blood-brain barrier (BBB) function of the co-culture model under normal condition. Then, we further determined the BBB function, inflammation, cell injury and the variation of neuroprotective factor in this model after anoxia-reoxygenation. The results suggest that this model exhibited a better BBB function and significantly increased expression of P-glycoprotein (Pg-P) and ZO-1 compared with BMECs only or co-culture with astrocytes or neurons. After anoxia-reoxygenation, the pathological changes of this model were basically resemblance to the pathological changes of brain cells and BBB in vivo. And nimodipine, an antagonist of calcium, could reverse those changes as well. According to our observations, we deduce that this triple cell co-culture model exhibits the basic structure, function and cell-cell interaction of NVU, which may offer a more proper in vitro system of NVU for the further investigation of cerebral diseases and drug screening.

Protection of the brain following cerebral ischemia through the attenuation of PARP-1-induced neurovascular unit damage in rats

“…The concept of the NVU was first proposed in 2003 (Lo et al, 2003), and it refers to an overall structure and functional unit that includes neurons, glial cells, endothelial cells, pericytes, basement membranes, microglia cells and the extracellular matrix. The NVU emphasizes the dynamic interactions among its components and their important influence on the pathophysiology of ischemia (Xue et al, 2013).…”

“…However, due to the narrow therapeutic window and hemorrhagic complications, only a minority of patients receive rt-PA therapy. Further analyses have documented that the key reason for these failures may be that only a single therapeutic target is pursued and that the interactions among the various components of the neurovascular unit (NVU) are neglected (Xue et al, 2013). Indeed, to obtain substantial and long-lasting neuroprotection, not only neurons but all of the components of the NVU, such as glia, endothelia, pericytes and basal membranes, should be targeted during an ischemic insult (Iadecola et al, 2006).…”

Setting-up an <em>In Vitro</em> Model of Rat Blood-brain Barrier (BBB): A Focus on BBB Impermeability and Receptor-mediated Transport

“…No "ideal" in vitro BBB model has been proposed yet 71,72,73 because the endothelial cell monolayers lack a number of important constituents of the neuro-glia vascular unit (NGVU) and are isolated from blood and body regulation. The lack of pericytes 74,16 or neurons 17,18 or the different constituents of the extracellular matrix used for plastic coating or the culture medium and serum used for cell growth in the most common and "easiest made" in vitro BBB models based on endothelial cells differentiated with astrocytes may modulate protein expression in comparison with the in vivo situation 75 . These models may express many transporters displayed in vivo, but not all.…”

Protection of Tong-Qiao-Huo-Xue Decoction against Cerebral Ischemic Injury through Reduction Blood–Brain Barrier Permeability

“…4,5) These layered cell structures construct the so-called tight junction (TJ) and the TJ-associated protein of Claudin-5, Occludin and ZO-1, which contribute to the integrity of the BBB, and the changes of its composition and the expression of relative ingredients are closely associated with BBB permeability. [6][7][8] These proteins are the major structural protein of TJ, their expressions levels are relevant to the degree of cerebral microvascular permeability and cerebral edema.…”