On-chip three-dimensional tumor spheroid formation and pump-less perfusion culture using gravity-driven cell aggregation and balanced droplet dispensing

Kim, Taeyoon; Doh, Il; Cho, Young-Ho

doi:10.1063/1.4739460

Cited by 28 publications

(26 citation statements)

References 26 publications

(28 reference statements)

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“…Moreover, imaging and biochemical analyses of the spheroids cannot be performed in situ under the above-mentioned culture conditions, requiring transfer of the spheroids to a different platform. To circumvent some of the issues mentioned above, Kim et al recently developed a well-based pump less perfusion culture system 14 . However none of these systems permit high throughput screening (HTS), which has been made possible using microfluidic methods for generation and processing of multicellular spheroids.…”

Section: Introductionmentioning

confidence: 99%

Generation and functional assessment of 3D multicellular spheroids in droplet based microfluidics platform

et al. 2016

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Here we describe a robust, microfluidic technique to generate and analyze 3D tumor spheroids, which resembles tumor microenvironment and can be used as a more effective preclinical drug testing and screening model. Monodisperse cell-laden alginate droplets were generated in Polydimethylsiloxane (PDMS) microfluidic devices that combine T-junction droplet generation and external gelation for spheroid formation. The proposed approach has the capability to incorporate multiple cell types. For the purposes of our study, we generated spheroids with breast cancer cell lines (MCF-7 drug sensitive and resistant) and co-culture spheroids of MCF-7 together with a fibroblast cell line (HS-5). The device has the capability to house 1000 spheroids on chip for drug screening and other functional analysis. Cellular viability of spheroids in the array part of the device was maintained for two weeks by continuous perfusion of complete media into the device. The functional performance of our 3D tumor models and a dose dependent response of standard chemotherapeutic drug, Doxorubicin (Dox) and standard drug combination Dox and Paclitaxel (PCT) was analyzed on our chip-based platform. Altogether, our work provides a simple and novel, in vitro platform to generate, image and analyze uniform, 3D monodisperse Alginate hydrogel tumors for various Omic studies and therapeutic efficiency screening, an important translational step before in vivo studies.

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Section: Introductionmentioning

confidence: 99%

Generation and functional assessment of 3D multicellular spheroids in droplet based microfluidics platform

et al. 2016

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“…Microfabricated microwell arrays and microfluidic devices have also been used for 3D cell culture. (17-19) These technologies are limited by expense and difficulties with exchange of media. More importantly, resulting spheroids are not individually addressable with drug compounds and downstream biochemical analyses reagents.…”

Section: Introductionmentioning

confidence: 99%

Optimization of Aqueous Biphasic Tumor Spheroid Microtechnology for Anti-cancer Drug Testing in 3D Culture

et al. 2014

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Tumor spheroids are three-dimensional clusters of cancer cells that exhibit characteristics of poorly perfused tumors and hence present a relevant model for testing the efficacy of anti-cancer compounds. The use of spheroids for drug screening is hindered by technological complexities for high throughput generation of consistent size spheroids individually addressable by drug compounds. Here we present and optimize a simple spheroid technology based on the use of an aqueous two-phase system. Cancer cells confined in a drop of the denser aqueous dextran phase are robotically dispensed into a microwell containing the immersion aqueous polyethylene glycol phase. Cells remain within the drop and form a viable spheroid, without a need for any external stimuli. The size of resulting spheroids is sensitive to volume variations of dispensed drops from the air displacement pipetting head of a commercial liquid handling robot. Therefore, we parametrically optimize the process of dispensing of dextran phase drops. For a given cell density, this optimization reproducibly generates consistent size spheroids in standard 96-well plates. In addition, we evaluate the use of a commercial biochemical assay to examine cellular viability of cancer cell spheroids. Spheroids show a dose-dependent response to cisplatin similar to a monolayer culture. However unlike their two-dimensional counterpart, spheroids exhibit resistance to paclitaxel treatment. This technology, which uses only commercially-available reagents and equipment, can potentially expedite anti-cancer drug discovery. Although the use of robotics makes the ATPS spheroid technology particularly useful for drug screening applications, this approach is compatible with simpler liquid handling techniques such as manual micropipetting and offers a straightforward method of 3D cell culture in research laboratories.

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“…The integration of 3D cell cultures with microfluidics technology can be used to create tumor models with perfusion to facilitate mass transport 16, 17 . Such microfluidic devices have been used to study various aspects of cancer progression such as tumor growth, presence of high interstitial fluid pressure, and cancer cell extravasation 18–24 .…”

Section: Introductionmentioning

confidence: 99%

Chemotaxis-driven assembly of endothelial barrier in a tumor-on-a-chip platform

et al. 2016

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The integration of three-dimensional micropatterning with microfluidics provides a unique opportunity to create perfusable tissue constructs in vitro. Herein, we have used this approach to create a tumor-on-chip with endothelial barrier. Specifically, we photopatterned a mixture of endothelial cells and cancer spheroids within a gelatin methacrylate (GelMA) hydrogel inside of a microfluidic device. The differential motility of endothelial and cancer cells in response to a controlled morphogen gradient across the cell-laden network drove the migration of endothelial cells to the periphery while maintaining the cancer cells within the interior of the hydrogel. The resultant endothelial cell layer forming cell-cell contact via VE-Cadherin junctions was found to encompass the entire the GelMA hydrogel structure. Furthermore, we have also examined the potential of such tumor-on-a-chip system as a drug screening platform using Doxorubicin, a model cancer drug.

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On-chip three-dimensional tumor spheroid formation and pump-less perfusion culture using gravity-driven cell aggregation and balanced droplet dispensing

Cited by 28 publications

References 26 publications

Generation and functional assessment of 3D multicellular spheroids in droplet based microfluidics platform

Generation and functional assessment of 3D multicellular spheroids in droplet based microfluidics platform

Optimization of Aqueous Biphasic Tumor Spheroid Microtechnology for Anti-cancer Drug Testing in 3D Culture

Chemotaxis-driven assembly of endothelial barrier in a tumor-on-a-chip platform

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