Transport‐mediated angiogenesis in 3D epithelial coculture

Sudo, Ryo; Chung, Seok; Zervantonakis, Ioannis K.; Vickerman, Vernella; Toshimitsu, Yasuko; Griffith, Linda G.; Kamm, Roger D.

doi:10.1096/fj.08-122820

Cited by 188 publications

(178 citation statements)

References 30 publications

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“…The microfluidic device design is based on previous work from our lab on forming endothelial sprouts under growth-factor gradients and the fabrication protocols are described in detail elsewhere (46). Compared with previous microfluidic systems from our group, two unique characteristics of the present design are (i) the incorporation of a y-junction for precise control of concentration gradients, required for accurate measures of endothelial permeability and (ii) the high number (n ¼ 37) of hydrogel ECM regions (Fig.…”

Section: Methodsmentioning

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.

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771

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: Methodsmentioning

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.

Self Cite

771

695

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“…1a). Channel width and height were selected to reduce shear stress on the cells during medium replacement and to provide sufficient area for cell adhesion to the sidewall of the gel scaffold (Sudo et al 2009). Details of the device fabrication process were described recently (Shin et al 2012).…”

Section: Preparation Of Microfluidic Devicesmentioning

confidence: 99%

“…Returning the device to horizontal, cells were then cultured under constant flow condition in order to promote cell adhesion to the gel. Interstitial flow was generated as described in our previous study (Sudo et al 2009). Briefly, medium reservoirs were inserted into the channel outlets (Fig.…”

Section: Protrusion Formation Assay and Interstitial Flow Generationmentioning

confidence: 99%

“…This microfluidic device has been used for hepatocyte-endothelial cell co-culture in our previous study (Sudo et al 2009). The microfluidic system has many advantages over the conventional methods used for tumor invasion studies such as culture inserts: abilities to monitor 3D cell migration, to create co-culture with defined cellular locations, and to culture tumor cells in controlled biochemical and biophysical microenvironments.…”

Section: Introductionmentioning

confidence: 99%

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A three-dimensional microfluidic tumor cell migration assay to screen the effect of anti-migratory drugs and interstitial flow

Kalchman

Fujioka

Chung³

et al. 2012

Microfluid Nanofluid

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Most anti-cancer drug screening assays are currently performed in two dimensions, on flat, rigid surfaces. However, there are increasing indications that threedimensional (3D) platforms provide a more realistic setting to investigate accurate morphology, growth, and sensitivity of tumor cells to chemical factors. Moreover, interstitial flow plays a pivotal role in tumor growth. Here, we present a microfluidic 3D platform to investigate behaviors of tumor cells in flow conditions with anti-migratory compounds. Our results show that interstitial flow and its direction have significant impact on migration and growth of hepatocellular carcinoma cell lines such as HepG2 and HLE. In particular, HepG2/HLE cells tend to migrate against interstitial flow, and their growth increases in interstitial flow conditions regardless of the flow direction. Furthermore, this migratory activity of HepG2 cells is enhanced when they are co-cultured with human umbilical vein endothelial cells. We also found that migration activity of HepG2 cells attenuates under hypoxic conditions. In addition, the effect of Artemisinin, an antimigratory compound, on HepG2 cells was quantitatively analyzed. The microfluidic 3D platform described here is useful to investigate more accurately the effect of antimigratory drugs on tumor cells and the critical influence of interstitial flow than 2D culture models.

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“…Roger Kamm's group have established 3D angiogenesis and capillary morphogenesis models, 190,191 where human endothelial cells extend their filopodial projections, migrating into the collagen matrix and forming open tube/lumen like structures when supplemented with proangiogenic factors. Angiogenesis was observed 192 when culturing rat hepatocytes and rat endothelial cells on each microfluidic side wells with a collagen gel scaffold in between. The rat endocytes formed capillary structures that extended into the hepatocytes channel and further fluorescent dextran protein diffused across the gel scaffold, demonstrating secreted proteins by either of the cell types.…”

Section: B Tissue Fabricationmentioning

confidence: 99%

Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics

Choudhury

Iliescu

et al. 2011

Biomicrofluidics

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There are a plethora of approaches to construct microtissues as building blocks for the repair and regeneration of larger and complex tissues. Here we focus on various physical and chemical trapping methods for engineering three-dimensional microtissue constructs in microfluidic systems that recapitulate the in vivo tissue microstructures and functions. Advances in these in vitro tissue models have enabled various applications, including drug screening, disease or injury models, and cellbased biosensors. The future would see strides toward the mesoscale control of even finer tissue microstructures and the scaling of various designs for high throughput applications. These tools and knowledge will establish the foundation for precision engineering of complex tissues of the internal organs for biomedical applications.

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Transport‐mediated angiogenesis in 3D epithelial coculture

Cited by 188 publications

References 30 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 three-dimensional microfluidic tumor cell migration assay to screen the effect of anti-migratory drugs and interstitial flow

Exploitation of physical and chemical constraints for three-dimensional microtissue construction in microfluidics

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