Sprouting Angiogenesis under a Chemical Gradient Regulated by Interactions with an Endothelial Monolayer in a Microfluidic Platform

Jeong, Gi Seok; Han, Sewoon; Shin, Yoojin; Kwon, Gu Han; Kamm, Roger D.; Lee, Sang Hoon; Chung, Seok

doi:10.1021/ac202170e

Cited by 105 publications

(87 citation statements)

References 42 publications

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“…[11][12][13][14][15] To verify that the device design with non-straight pillars allows the formation of a concentration gradient within the central channel, we performed preliminary numerical simulations in COMSOL. Generally, the generation and maintenance of a stable concentration gradient between the source and the sink depend on the diffusion coefficient of the diffusing molecule within the hydrogel, as well as on the geometrical features of the device.…”

Section: Resultsmentioning

confidence: 99%

“…Different devices based on this approach have been developed in order to study and unravel the intricate cell-signal interactions that occur in vivo. [13][14][15] Although these lFds may differ in terms of shape, dimensions, and culturing conditions, they all rely on the resistance effects of a cell laden biopolymeric gel to the flow of soluble molecules of interest. The presence of short cuts within the cell containing 3D matrix affects the spatial distribution of molecules and matrix collapse or the local densification impairs the diffusion mechanisms.…”

Section: Introductionmentioning

confidence: 99%

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Optimizing design and fabrication of microfluidic devices for cell cultures: An effective approach to control cell microenvironment in three dimensions

et al. 2014

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The effects of gradients of bioactive molecules on the cell microenvironment are crucial in several biological processes, such as chemotaxis, angiogenesis, and tumor progression. The elucidation of the basic mechanisms regulating cell responses to gradients requires a tight control of the spatio-temporal features of such gradients. Microfluidics integrating 3D gels are useful tools to fulfill this requirement. However, even tiny flaws in the design or in the fabrication process may severely impair microenvironmental control, thus leading to inconsistent results. Here, we report a sequence of actions aimed at the design and fabrication of a reliable and robust microfluidic device integrated with collagen gel for cell culturing in 3D, subjected to a predetermined gradient of biomolecular signals. In particular, we developed a simple and effective solution to the frequently occurring technical problems of gas bubble formation and 3D matrix collapsing or detaching from the walls. The device here proposed, in Polydimethylsiloxane, was designed to improve the stability of the cell-laden hydrogel, where bubble deprived conditioning media flow laterally to the gel. We report the correct procedure to fill the device with the cell populated gel avoiding the entrapment of gas bubbles, yet maintaining cell viability. Numerical simulations and experiments with fluorescent probes demonstrated the establishment and stability of a concentration gradient across the gel. Finally, chemotaxis experiments of human Mesenchymal Stem Cells under the effects of Bone Morphogenetic Protein-2 gradients were performed in order to demonstrate the efficacy of the system in controlling cell microenvironment. The proposed procedure is sufficiently versatile and simple to be used also for different device geometries or experimental setups

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Optimizing design and fabrication of microfluidic devices for cell cultures: An effective approach to control cell microenvironment in three dimensions

et al. 2014

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“…Human MSCs were isolated from fresh bone marrow aspirates obtained from Cambrex Life Sciences as previously described (38,50), grown in high-glucose DMEM with 10% (vol/vol) FBS and 1% penicillin/streptomycin (all from Invitrogen) with 0.1 ng/mL basic FGF (Invitrogen), and expanded to passage 8 or lower. Human umbilical vein endothelial cells were isolated from postnatal discarded umbilical veins and fully de-identified following the protocol IRB-AAAC4839 approved by the Columbia University Institutional Review Board, and grown using EGM-2 Media (Lonza) to passage 8 or lower.…”

Section: Methodsmentioning

confidence: 99%

“…For example, endothelial cell (EC) stabilization via mesenchymal stem cell (MSC) interactions is known to facilitate the maturation of blood vessels impacting many physiologic systems, from tumors to engineered tissues (33)(34)(35). Recent studies clearly showed the importance of microscale gradients for vasculogenesis (36)(37)(38).…”

mentioning

confidence: 99%

Assembly of complex cell microenvironments using geometrically docked hydrogel shapes

Eng

Lee

Parsa³

et al. 2013

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

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Cellular communities in living tissues act in concert to establish intricate microenvironments, with complexity difficult to recapitulate in vitro. We report a method for docking numerous cellularized hydrogel shapes (100-1,000 μm in size) into hydrogel templates to construct 3D cellular microenvironments. Each shape can be uniquely designed to contain customizable concentrations of cells and molecular species, and can be placed into any spatial configuration, providing extensive compositional and geometric tunability of shapecoded patterns using a highly biocompatible hydrogel material. Using precisely arranged hydrogel shapes, we investigated migratory patterns of human mesenchymal stem cells and endothelial cells. We then developed a finite element gradient model predicting chemotactic directions of cell migration in micropatterned cocultures that were validated by tracking ∼2,500 individual cell trajectories. This simple yet robust hydrogel platform provides a comprehensive approach to the assembly of 3D cell environments.microtechnologies | tissue assembly | angiogenesis | modeling | diffusion B iological tissues are composed of cellular "building blocks" that cooperate to provide tissue-specific functions (1-6). Specific cells, molecules, and their geometric assembly establish a biological system, whether it is a vascular network surrounded by parenchymal cells (7), a developing tissue (8-11), or a metastatic tumor (12-16). In vitro culture systems designed to control the 3D presentation of multiple cells and molecular species in a biologically relevant matrix are needed to faithfully recapitulate intricate biological niches (17-23). Previous attempts to assemble complex tissue structures in vitro lacked the specificity and yield to control large numbers of cells and soluble factors (22-24), had limited resolution at the microscale (25, 26), and used matrices without adequate biological function (27-32). Therefore, spatial microenvironmental control of biological systems has been difficult to achieve. Such control is important in many processes, including the migratory formation of vasculature, where gradient patterns dictate growth of vascular sprouts. For example, endothelial cell (EC) stabilization via mesenchymal stem cell (MSC) interactions is known to facilitate the maturation of blood vessels impacting many physiologic systems, from tumors to engineered tissues (33-35). Recent studies clearly showed the importance of microscale gradients for vasculogenesis (36-38).We report a method by which microsized 3D hydrogels are shape-coded for their biological and physical properties and docked by iterative sedimentation into shape-matching hydrogel templates. Microenvironmental niches were fabricated using gelatin methacrylate (GelMA), a modified native protein with excellent biocompatibility, tunable mechanical properties, and micrometer-scale patterning resolution (39, 40). GelMA was molded into diverse geometric shapes with dimensions of 100-1,000 μm, after encapsulating cells or labeled molecular specie...

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“…23 Researchers have taken advantage of these microfluidic platforms and successfully developed in vitro models for studying various important physiological phenomena, such as wound healing, 24,25 cancer metastasis, 26,27 and vascular angiogenesis. 28,29 Among these innovative microfluidic devices, some are dedicated for co-culture system, which is a rather challenging task compared to handling a single type of cells on a chip. Prior reports have shown various designs, including valves, 30 laminar flow between immiscible liquids, 31,32 separate compartments for dynamic co-culture, 33 and detachable substrates, where the cells are individually cultured on different substrates.…”

Section: Introductionmentioning

confidence: 99%

A microfluidic co-culture system to monitor tumor-stromal interactions on a chip

et al. 2014

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The living cells are arranged in a complex natural environment wherein they interact with extracellular matrix and other neighboring cells. Cell-cell interactions, especially those between distinct phenotypes, have attracted particular interest due to the significant physiological relevance they can reveal for both fundamental and applied biomedical research. To study cell-cell interactions, it is necessary to develop co-culture systems, where different cell types can be cultured within the same confined space. Although the current advancement in lab-on-a-chip technology has allowed the creation of in vitro models to mimic the complexity of in vivo environment, it is still rather challenging to create such co-culture systems for easy control of different colonies of cells. In this paper, we have demonstrated a straightforward method for the development of an on-chip co-culture system. It involves a series of steps to selectively change the surface property for discriminative cell seeding and to induce cellular interaction in a co-culture region. Bone marrow stromal cells (HS5) and a liver tumor cell line (HuH7) have been used to demonstrate this co-culture model. The cell migration and cellular interaction have been analyzed using microscopy and biochemical assays. This co-culture system could be used as a disease model to obtain biological insight of pathological progression, as well as a tool to evaluate the efficacy of different drugs for pharmaceutical studies. V C 2014 AIP Publishing LLC. [http://dx

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Sprouting Angiogenesis under a Chemical Gradient Regulated by Interactions with an Endothelial Monolayer in a Microfluidic Platform

Cited by 105 publications

References 42 publications

Optimizing design and fabrication of microfluidic devices for cell cultures: An effective approach to control cell microenvironment in three dimensions

Optimizing design and fabrication of microfluidic devices for cell cultures: An effective approach to control cell microenvironment in three dimensions

Assembly of complex cell microenvironments using geometrically docked hydrogel shapes

A microfluidic co-culture system to monitor tumor-stromal interactions on a chip

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