Permeability characteristics of cultured endothelial cell monolayers

Albelda, Steven Μ.; Sampson, Phyllis; Haselton, Frederick R.; McNiff, Jennifer M.; Mueller, Stephen N.; Williams, Stuart K.; Fishman, Alfred P.; Levine, Elliot M.

doi:10.1152/jappl.1988.64.1.308

Cited by 185 publications

(129 citation statements)

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“…Endothelial barrier function quantitation in the presence of invading cells showed size-selective transendothelial transport and the P D values in our microfluidic model agree well with measurements in transwell systems (37) and in engineered blood vessels in 3D matrices (38). Although our measurements are significantly higher than in vivo values of healthy vasculature (39), they are of the same order of magnitude ( Fig.…”

Section: Discussionsupporting

confidence: 84%

Three-dimensional microfluidic model for tumor cell intravasation and endothelial barrier function

Zervantonakis

Hughes

Charest

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

770

695

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Entry of tumor cells into the blood stream is a critical step in cancer metastasis. Although significant progress has been made in visualizing tumor cell motility in vivo, the underlying mechanism of cancer cell intravasation remains largely unknown. We developed a microfluidic-based assay to recreate the tumor-vascular interface in three-dimensions, allowing for high resolution, real-time imaging, and precise quantification of endothelial barrier function. Studies are aimed at testing the hypothesis that carcinoma cell intravasation is regulated by biochemical factors from the interacting cells and cellular interactions with macrophages. We developed a method to measure spatially resolved endothelial permeability and show that signaling with macrophages via secretion of tumor necrosis factor alpha results in endothelial barrier impairment. Under these conditions intravasation rates were increased as validated with live imaging. To further investigate tumor-endothelial (TC-EC) signaling, we used highly invasive fibrosarcoma cells and quantified tumor cell migration dynamics and TC-EC interactions under control and perturbed (with tumor necrosis factor alpha) barrier conditions. We found that endothelial barrier impairment was associated with a higher number and faster dynamics of TC-EC interactions, in agreement with our carcinoma intravasation results. Taken together our results provide evidence that the endothelium poses a barrier to tumor cell intravasation that can be regulated by factors present in the tumor microenvironment.T umor-endothelial cell interactions are critical in multiple steps during cancer metastasis, ranging from cancer angiogenesis to colonization. Cancer cell intravasation is a rate-limiting step in metastasis that regulates the number of circulating tumor cells and thus presents high risk for the formation of secondary tumors (1, 2). During the metastatic process tumor cells migrate out of the primary tumor (3), navigate into a complex tumor microenvironment, and enter into blood vessels (4). Cell-cell communication and chemotaxis (5) are key to this process and can occur via paracrine signals and/or direct contact between different cell types during tumor cell invasion (6) and metastatic colonization (7). Studies using multiphoton imaging in animal models have demonstrated that the ability of tumor cells to enter into the blood stream can be controlled both by tumor cell intrinsic factors (8-11) and other cells present in the tumor microenvironment, such as macrophages (12) and neutrophils (13). However, because of the lack of physiologically relevant in vitro models and the challenges of investigating cell-cell interactions in vivo, the underlying mechanism of intravasation remains poorly understood (14). In particular, a number of fundamental questions remain as to whether intravasation is an active or passive process (15) and whether tumor cells cross the endothelial barrier through cell-cell junctions (paracellular) or through the endothelial cell body [transcellular (16)]. Therefor...

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Section: Discussionsupporting

confidence: 84%

Three-dimensional microfluidic model for tumor cell intravasation and endothelial barrier function

Zervantonakis

Hughes

Charest

et al. 2012

Proc. Natl. Acad. Sci. U.S.A.

770

695

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“…8). It should be noted that it is not appropriate to compare a value of permeability obtained from in vitro experiments to that of the in vivo experiments because EC permeability of in vitro system is 10 to 100 times greater than that of intact continuous endothelium (24) . However, in vitro system has an advantage to make clear a response of ECs to mechanical environments.…”

Section: Discussionmentioning

confidence: 99%

Effect of Shear Stress on Permeability of Vascular Endothelial Monolayer Cocultured with Smooth Muscle Cells

Sakamoto

Ohashi

Sato

2004

JSME Int. J., Ser. C

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Effect of fluid shear stress on permeability of endothelial monolayer was investigated using an endothelial cell (EC)-smooth muscle cell (SMC) cocultured model (CM). Permeability of ECs to bovine serum albumin was measured after exposure to shear stress of 1.5 Pa for 48 hours. Morphology and VE-cadherin expression of ECs in CM was almost same as of ECs cultured alone (monocultured model, MM). Under static condition, EC permeability was 5.1 ± 3.0 × 10 −6 cm/sec (mean ± SD) in MM and 6.5 ± 3.4 × 10 −6 cm/sec in CM. After exposure to shear stress, EC permeability in CM (2.2 ± 1.9 × 10 −6 cm/sec, p < 0.05) significantly decreased compared with the static model. However, EC permeability in MM (3.9±3.2×10 −6 cm/sec) did not significantly change compared with static cultured condition. These results suggested that cellular interactions between ECs and SMCs have important influences on EC permeability.

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“…Fluorescence in the 24-well plates was measured (excitation at 485 nm; emission at 535 nm) in a Tecan SPECTRAfluor microplate reader. BSA diffusional permeability was calculated from the ratio of the measured fluorescence concentrations in the top and bottom wells [17].…”

Section: Methodsmentioning

confidence: 99%

Effect of advanced glycation end products on oxidative stress in endothelial cells in culture: a warning on the use of cells studied in serum-free media

et al. 2001

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A common cardiovascular complication of diabetes is an increase in the vascular escape rate of albumin, generally interpreted as an increase in microvascular permeability [1±3]. One of the more recent theories to explain this endothelial dysfunction focuses on the accumulation in blood and tissues of the advanced glycation end products (AGEs) formed from protein modification by glucose [4]. The AGE hypothesis holds that chronic diabetes-accelerated chemical modification of proteins, lipids and other molecules by reducing sugars alters their structure and function, inducing cellular Diabetologia (2001) Abstract Aims/hypothesis. Alterations in vascular permeability and oxidative stress are characteristics of endothelial dysfunction in diabetic vascular disease. Since AGE-proteins have been hypothesized to mediate these effects, we studied the effects of AGE-bovine serum albumin on endothelial monolayer permeability and intracellular glutathione. Methods. AGE-BSA was prepared by incubating BSA for 30 days at 37 C with 0.5 mol/l glucose and 0.2 mol/l phosphate buffer, pH 7.4. Permeability to fluorescently labelled BSA was assessed in a bovine pulmonary artery endothelial cell monolayer preparation. Glutathione was measured by an enzymatic assay. Results. AGE-BSA concentrations greater than 3 to 4 mmol/l produced maximal increases in permeability (6±8 times basal) within 3 to 4 h of incubation with the cells. This effect persisted for at least 48 h. However, BSA incubated in the absence of glucose produced similar effects. Dialysis of the AGE-BSA showed that low molecular weight components contained the permeability-increasing activity. Phosphate buffer used to prepare the AGE-BSA, at concentrations equivalent to those present in phosphate-buffered saline and in the AGE preparation (~5 mmol/l), produced similar permeability increases at equivalent incubation times. Metal chelators (0.5 mmol/l) or inclusion of fetal bovine serum (10±20 %) blocked these permeability increases. These increases in permeability were associated with a decrease in endothelial glutathione, both inhibited by 10 mmol/l N-acetylcysteine, and a loss of cell-tocell and cell-to-matrix adhesion molecules. Conclusion/interpretation. Trace amounts of redoxactive metal ions in biological buffers could induce oxidative stress and alterations in cellular functions attributed to AGE-proteins in vitro. It is important to use metal-free phosphate and bicarbonate buffers in studies on cell biology in vitro, especially in serum-free media. [Diabetologia (2001

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Permeability characteristics of cultured endothelial cell monolayers

Cited by 185 publications

References 0 publications

Three-dimensional microfluidic model for tumor cell intravasation and endothelial barrier function

Three-dimensional microfluidic model for tumor cell intravasation and endothelial barrier function

Effect of Shear Stress on Permeability of Vascular Endothelial Monolayer Cocultured with Smooth Muscle Cells

Effect of advanced glycation end products on oxidative stress in endothelial cells in culture: a warning on the use of cells studied in serum-free media

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