Microtissue size and hypoxia in HTS with 3D cultures

Asthana, Amish; Kisaalita, William S.

doi:10.1016/j.drudis.2012.03.004

Cited by 81 publications

(64 citation statements)

References 62 publications

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“…Culturing cells in a monolayer has certain advantages; the cells are simple to handle and image, and one can rely on established techniques and standard laboratory equipment. Two-dimensional cell cultures, however, have certain limitations in mimicking functional living tissue, as original microenvironmental inputs, such as cell-to-cell interaction and spatiotemporal biochemical gradients, are absent [7][8][9] . Three-dimensional (3D) cell cultures offer the potential to overcome many of these limitations and are therefore finding increasing applications in the pharmaceutical industry and basic research 10 .…”

Section: Introductionmentioning

confidence: 99%

Multi-analyte biosensor interface for real-time monitoring of 3D microtissue spheroids in hanging-drop networks

et al. 2016

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Microfluidics is becoming a technology of growing interest for building microphysiological systems with integrated read-out functionalities. Here we present the integration of enzyme-based multi-analyte biosensors into a multi-tissue culture platform for 'body-on-a-chip' applications. The microfluidic platform is based on the technology of hanging-drop networks, which is designed for the formation, cultivation, and analysis of fluidically interconnected organotypic spherical three-dimensional (3D) microtissues of multiple cell types. The sensor modules were designed as small glass plug-ins featuring four platinum working electrodes, a platinum counter electrode, and an Ag/AgCl reference electrode. They were placed directly into the ceiling substrate from which the hanging drops that host the spheroid cultures are suspended. The electrodes were functionalized with oxidase enzymes to enable continuous monitoring of lactate and glucose through amperometry. The biosensors featured high sensitivities of 322 ± 41 nA mM − 1 mm − 2 for glucose and 443 ± 37 nA mM − 1 mm − 2 for lactate; the corresponding limits of detection were below 10 μM. The proposed technology enabled tissue-size-dependent, real-time detection of lactate secretion from single human colon cancer microtissues cultured in the hanging drops. Furthermore, glucose consumption and lactate secretion were monitored in parallel, and the impact of different culture conditions on the metabolism of cancer microtissues was recorded in real-time.

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Section: Introductionmentioning

confidence: 99%

Multi-analyte biosensor interface for real-time monitoring of 3D microtissue spheroids in hanging-drop networks

et al. 2016

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“…[6][7][8] Thus, compared to a monolayer, a multicellular tumor spheroid can better mimic aspects of the tumor in vivo including cell-cell contacts, 9 diffusion gradients of oxygen 10 and drugs 11 (when the spheroid is of sufficient size, typically several hundred micrometers in diameter [12][13][14], and cell-cell adhesion. 15 Previous work has found that heterogeneous spheroid sizes can give rise to different levels of hypoxia and thus different gene expression, 16 highlighting the need for good uniformity in spheroid size 17 within an assay. Recent methods to rapidly generate large numbers of uniform spheroids include microfabrication to control numbers of cells sedimenting into wells and thus the resultant spheroid size, 4 robotic dispensing of cells into molded hydrogel templates, 18 and hanging drop arrays.…”

Section: Introductionmentioning

confidence: 99%

Core-shell hydrogel beads with extracellular matrix for tumor spheroid formation

Grist

Nasseri

et al. 2015

Biomicrofluidics

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Creating multicellular tumor spheroids is critical for characterizing anticancer treatments since they may provide a better model of the tumor than conventional monolayer culture. Moreover, tumor cell interaction with the extracellular matrix can determine cell organization and behavior. In this work, a microfluidic system was used to form cell-laden core-shell beads which incorporate elements of the extracellular matrix and support the formation of multicellular spheroids. The bead core (comprising a mixture of alginate, collagen, and reconstituted basement membrane, with gelation by temperature control) and shell (comprising alginate hydrogel, with gelation by ionic crosslinking) were simultaneously formed through flow focusing using a cooled flow path into the microfluidic chip. During droplet gelation, the alginate acts as a fast-gelling shell which aids in preventing droplet coalescence and in maintaining spherical droplet geometry during the slower gelation of the collagen and reconstituted basement membrane components as the beads warm up. After droplet gelation, the encapsulated MCF-7 cells proliferated to form uniform spheroids when the beads contained all three components: alginate, collagen, and reconstituted basement membrane. The dose-dependent response of the MCF-7 cell tumor spheroids to two anticancer drugs, docetaxel and tamoxifen, was compared to conventional monolayer culture.

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“…This loss of function in 2D motivates the strong current effort to develop three-dimensional tissue structures, as they have been proven to exhibit properties that more closely resemble those of a living organism 6,[8][9][10] . In analogy to the 2D case, microfluidic systems can be used to better control culturing conditions and to render those as similar as possible to the in vivo situation.…”

mentioning

confidence: 99%

Reconfigurable microfluidic hanging drop network for multi-tissue interaction and analysis

Frey

Misun

Fluri³

et al. 2014

Nat Commun

335

303

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Integration of multiple three-dimensional microtissues into microfluidic networks enables new insights in how different organs or tissues of an organism interact. Here, we present a platform that extends the hanging-drop technology, used for multi-cellular spheroid formation, to multifunctional complex microfluidic networks. Engineered as completely open, 'hanging' microfluidic system at the bottom of a substrate, the platform features high flexibility in microtissue arrangements and interconnections, while fabrication is simple and operation robust. Multiple spheroids of different cell types are formed in parallel on the same platform; the different tissues are then connected in physiological order for multi-tissue experiments through reconfiguration of the fluidic network. Liquid flow is precisely controlled through the hanging drops, which enable nutrient supply, substance dosage and inter-organ metabolic communication. The possibility to perform parallelized microtissue formation on the same chip that is subsequently used for complex multi-tissue experiments renders the developed platform a promising technology for 'body-on-a-chip'-related research.

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Microtissue size and hypoxia in HTS with 3D cultures

Cited by 81 publications

References 62 publications

Multi-analyte biosensor interface for real-time monitoring of 3D microtissue spheroids in hanging-drop networks

Multi-analyte biosensor interface for real-time monitoring of 3D microtissue spheroids in hanging-drop networks

Core-shell hydrogel beads with extracellular matrix for tumor spheroid formation

Reconfigurable microfluidic hanging drop network for multi-tissue interaction and analysis

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