Generation of oxygen gradients in microfluidic devices for cell culture using spatially confined chemical reactions

Chen, Yung-Ann; King, Andrew; Shih, Hsiu-Chen; Peng, Chien-Chung; Wu, Chueh‐Yu; Liao, Wei-Hao; Tung, Yi-Chung

doi:10.1039/c1lc20325h

Cited by 162 publications

(178 citation statements)

References 24 publications

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“…The concentration gradient of small molecules can be controlled temporally and spatially (at length scales of tens of microns, which is the scale of a single cell) in a microfluidic device (7,8,(24)(25)(26)(27)(28). These devices are often fabricated in optically transparent materials and make it possible to visualize cell migration in real time (19).…”

Section: Introductionmentioning

confidence: 99%

A paper-based invasion assay: Assessing chemotaxis of cancer cells in gradients of oxygen

et al. 2015

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This work describes a 3D, paper-based assay that can isolate subpopulations of cells based on their invasiveness (i.e., distance migrated in a hydrogel) in a gradient of concentration of oxygen (O 2 ). Layers of paper impregnated with a cell-compatible hydrogel are stacked and placed in a plastic holder to form the invasion assay. Stacking the layers of paper assembles them into 3D tissue-like constructs of defined thickness and composition. The plastic holder ensures the layers of paper are in conformal contact; this geometry allows the cells to migrate between adjacent layers through the embedded hydrogel. In most assays, the stack comprises a single layer of paper containing mammalian cells suspended in a hydrogel, sandwiched between multiple layers of paper containing only hydrogel (into which the cells migrate). Cells in the stack consume and produce small molecules; these molecules diffuse throughout the stack to generate gradients both in the stack, and between the stack and the bulk culture medium. Placing the cell-containing layer in different positions of the stack, or modifying the permeability of the holder to oxygen or proteins, alters the profile of the gradients within the stack. Physically separating the layers after culture isolates subpopulations of cells that migrated different distances, and enables their subsequent analysis or culture. Using this system, three independent cell lines derived from A549 cancer cells are shown to produce distinguishable migration behavior in a gradient of oxygen. This result is the first experimental demonstration that oxygen acts as a chemoattractant for cancer cells. Page 3 of 35 Significance StatementThe invasion of cancerous cells from a tumor into surrounding tissues is one contribution to metastasis-a major contributor to death for patients with cancer. There is a strong link between the directed invasion of cancer cells and the gradients of molecules formed in the microenvironment of the tumor. Using a paper-based invasion assay, this work demonstrates that oxygen-a nutrient known to induce significant behavioral changes to cells within a tumor in a concentration-dependent manner-can also act as a chemoattractant, resulting in the migration of cancer cells towards higher concentrations of oxygen. This finding, and the invasion assay described, could lead to a better understanding of oxygen-based chemotaxis in cancer, and ultimately new strategies for managing metastasis.

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Section: Introductionmentioning

confidence: 99%

A paper-based invasion assay: Assessing chemotaxis of cancer cells in gradients of oxygen

et al. 2015

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“…[11][12][13][14][15][16][17] Microfluidics is capable of better controlling flows in spatial and temporal domains, which allows reconstituting precise and more in vivo-like microenvironments to study cell behaviors. Using microfluidic devices, cell spheroids can be cultured in a normal incubator and a particular spheroid can be monitored for certain periods of time.…”

Section: Introductionmentioning

confidence: 99%

A microfluidic device for uniform-sized cell spheroids formation, culture, harvesting and flow cytometry analysis

Patra¹,

Chen

Peng

et al. 2013

Biomicrofluidics

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Culture of cells as three-dimensional (3D) aggregates, named spheroids, possesses great potential to improve in vitro cell models for basic biomedical research. However, such cell spheroid models are often complicated, cumbersome, and expensive compared to conventional Petri-dish cell cultures. In this work, we developed a simple microfluidic device for cell spheroid formation, culture, and harvesting. Using this device, cells could form uniformly sized spheroids due to strong cell-cell interactions and the spatial confinement of microfluidic culture chambers. We demonstrated cell spheroid formation and culture in the designed devices using embryonic stem cells, carcinoma cells, and fibroblasts. We further scaled up the device capable of simultaneously forming and culturing 5000 spheroids in a single chip. Finally, we demonstrated harvesting of the cultured spheroids from the device with a simple setup. The harvested spheroids possess great integrity, and the cells can be exploited for further flow cytometry assays due to the ample cell numbers. V C 2013 AIP Publishing LLC. [http://dx

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“…Microfluid ic devices for creating a concentration gradient of a reagent and protein [8] and o xygen gradient [9,10] have been developed, and various microdevices for cell culture to facilitate microscopic observation of cellular behavior [11,12] and drug research [13,14] have been reported; some devices are available fo r cell cultures with an o xygen gradient [15,16]. However, the absence of a technique for co-cultures in microdevices to study interactions between cells of different types, has delayed the understanding of mechanis ms that relate to tumor metastasis under hypoxic conditions.…”

Section: Introductionmentioning

confidence: 99%

Co-culture Microdevice with Oxygen Gradient for Tumor Microenvironment Model and Metastasis Imaging

T¹,

H²,

Tsukada³

2012

AJBE

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Tu mor hypoxia is a major therapeutic problem since it decreases radiation effects and leads to metastasis.Oxygen is delivered to tumor tissue via abnormal and dysfunctional microvessels, which forms heterogeneity of tissue oxygenation in the tumo r. Mimicking the o xygen gradient fo r cellu lar experiments in vitro is important to clarify the mechanis ms involved in tumor bio logy, but the only method to produce hypoxic conditions at a constant level is using gas-controlled incubators, because there is currently no technique for creating an o xygen gradient using culture dishes. We designed a polydimethylsilo xane (PDMS) microflu idic device integrated with microchannels for cell cultures that enables visualizat ion of cellu lar distribution under a microscope and co -culture to determine interactions between cancer and other cells. Phosphorescence-based partial o xygen measurements quantified the o xygen grad ient, which can be controlled by the gas pressure between the inlet and outlet of the device. A monoculture of end othelial cells with an oxygen gradient in the device showed an increase in cell death in the hypoxic area. In addition, Lewis lung carcino ma cells co -cultured with endothelial cells showed gradient-dependent migration through a membrane pore filter, indicating that the interaction between tumor and endothelial cells under hypo xia is crucial in metastasis. The results suggest that the developed microdevice can be used to study the mechanisms of tu mor metastasis under hypoxic conditions.

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Generation of oxygen gradients in microfluidic devices for cell culture using spatially confined chemical reactions

Cited by 162 publications

References 24 publications

A paper-based invasion assay: Assessing chemotaxis of cancer cells in gradients of oxygen

A paper-based invasion assay: Assessing chemotaxis of cancer cells in gradients of oxygen

A microfluidic device for uniform-sized cell spheroids formation, culture, harvesting and flow cytometry analysis

Co-culture Microdevice with Oxygen Gradient for Tumor Microenvironment Model and Metastasis Imaging

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