Microfluidic Endothelium for Studying the Intravascular Adhesion of Metastatic Breast Cancer Cells

Song, Jonathan W.; Cavnar, Stephen P.; Walker, Ann C.; Luker, Kathryn E.; Gupta, Mudit; Tung, Yi-Chung; Takayama, Shuichi

doi:10.1371/journal.pone.0005756

Cited by 297 publications

(286 citation statements)

References 52 publications

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“…For example, a recent in vitro study identified changes in endothelial myosin light chain kinase upon tumor-endothelial cell contact (16), however this method did not allow for measurement of endothelial permeability or the accurate control of the microenvironmental stimuli. Compared to other in vitro models (16,21) and microfluidic models of tumor-endothelial interactions (25,35), our assay provides a number of unique features, such as high-resolution live cell imaging, the formation of an endothelial monolayer on a 3D ECM enabling us to model intravasation, and for precise quantification and control of critical tumor microenvironmental factors. The readily accessible apical side of the endothelium allows for the introduction of cell types in the tumor microenvironment, such as macrophages, the precise establishment of growth-factor gradients, fluid flow, and real-time spatially resolved endothelial barrier function measurements.…”

Section: Discussionmentioning

confidence: 99%

“…Hence, modeling the tumor microenvironment by integrating the interactions among multiple cell types with biochemical and biophysical factors is a very attractive target for microfluidics. Toward this end, a number of microfluidic designs have been developed to study growth-factor gradients in cancer cell migration (22,23), tumor-stromal cell interactions (24), and tumor-endothelial cell interactions (25).…”

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confidence: 99%

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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: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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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“…Others have detailed the process by which porous membranes are incorporated into microdevices 7,12,23 and demonstrated their utility for biological studies. 7,8,12,24,25 For the devices used in this study, the top straight channel and bottom S-shaped channel were 2000 m wide by 300 m high.…”

Section: A Device Fabricationmentioning

confidence: 99%

A microfluidic membrane device to mimic critical components of the vascular microenvironment

Srigunapalan¹,

Lam²,

Wheeler³

et al. 2011

Biomicrofluidics

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Vascular function, homeostasis, and pathological development are regulated by the endothelial cells that line blood vessels. Endothelial function is influenced by the integrated effects of multiple factors, including hemodynamic conditions, soluble and insoluble biochemical signals, and interactions with other cell types. Here, we present a membrane microfluidic device that recapitulates key components of the vascular microenvironment, including hemodynamic shear stress, circulating cytokines, extracellular matrix proteins, and multiple interacting cells. The utility of the device was demonstrated by measuring monocyte adhesion to and transmigration through a porcine aortic endothelial cell monolayer. Endothelial cells grown in the membrane microchannels and subjected to 20 dynes/ cm 2 shear stress remained viable, attached, and confluent for several days. Consistent with the data from macroscale systems, 25 ng/ml tumor necrosis factor ͑TNF͒-␣ significantly increased RAW264.7 monocyte adhesion. Preconditioning endothelial cells for 24 h under static or 20 dynes/ cm 2 shear stress conditions did not influence TNF-␣-induced monocyte attachment. In contrast, simultaneous application of TNF-␣ and 20 dynes/ cm 2 shear stress caused increased monocyte adhesion compared with endothelial cells treated with TNF-␣ under static conditions. THP-1 monocytic cells migrated across an activated endothelium, with increased diapedesis in response to monocyte chemoattractant protein ͑MCP͒-1 in the lower channel of the device. This microfluidic platform can be used to study complex cell-matrix and cell-cell interactions in environments that mimic those in native and tissue engineered blood vessels, and offers the potential for parallelization and increased throughput over conventional macroscale systems.

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“…These models have been used to study the interactions between the endothelium and other cells, such as leukocytes or cancer cells. [30][31][32][33][34][35][36] However, most current microfluidic blood vessel models only provide a controlled environment for adhesion of individual cancer cells to the endothelium under stimuli. Limited information is available regarding the interaction of tumor aggregates and endothelium, as well as the subsequent events (such as transendothelial invasion or proliferation in situ) that follow adhesion.…”

Section: Introductionmentioning

confidence: 99%

A microfluidic-based device for study of transendothelial invasion of tumor aggregates in realtime

2012

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Circulating tumor aggregates exhibit a high metastatic potential and could potentially serve as an important target for cancer therapies. In this study, we developed a microfluidic model that reconstitutes and is representative of the principal components of biological blood vessels, including vessel cavity, endothelium, and perivascular matrix containing chemokines. Using this model, the transendothelial invasion of tumor aggregates can be observed and recorded in realtime. In this study we analyzed the extravasation process of salivary gland adenoid cystic carcinoma (ACC) cell aggregates. ACC aggregates transmigrated across the endothelium under the stimulation of chemokine CXCL12. The endothelial integrity was irreversibly damaged at the site of transendothelial invasion. The transendothelial invasion of ACC aggregates was inhibited by AMD3100, but the adhesion of ACC aggregates to the endothelium was not affected by the CXCR4 antagonist. This model allows for detailed study of the attachment and transendothelial invasion of tumor aggregates; thus, it would be a useful tool for analysis of the underlying mechanisms of metastasis and for testing novel anti-metastasis agents.

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Microfluidic Endothelium for Studying the Intravascular Adhesion of Metastatic Breast Cancer Cells

Cited by 297 publications

References 52 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

A microfluidic membrane device to mimic critical components of the vascular microenvironment

A microfluidic-based device for study of transendothelial invasion of tumor aggregates in realtime

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