A multiscale, vertical-flow perfusion system with integrated porous microchambers for upgrading multicellular spheroid culture

Takagi, M.; Yamada, Masao; Utoh, Rie; Seki, Masayuki

doi:10.1039/d3lc00168g

Cited by 6 publications

(5 citation statements)

References 65 publications

(86 reference statements)

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“…The vertical-flow perfusion through the pores improved cell functions because the flow provided oxygen and nutrients to the spheroids. 35 The porous membrane electrodes can evaluate cell functions while maintaining the cell culture environment. In addition, porous membrane electrodes have the potential for evaluating not only spheroids but also organoids in MPSs through glucose uptake measurements for drug screening.…”

Section: ■ Results and Discussionmentioning

confidence: 99%

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Enzyme-Free In-Situ Electrochemical Measurement Using a Porous Membrane Electrode for Glucose Transport into Cell Spheroids

Utagawa,

Ino,

Shinoda

et al. 2024

ACS Sens.

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Microphysiological systems have attracted attention because of their use in drug screening. However, it is challenging to measure cell functions in real time using a device. In this study, we developed a cell culture device using a porous membrane electrode for in situ electrochemical glucose measurements for cell analysis. First, a porous membrane electrode was fabricated and electrochemically evaluated for enzymefree glucose measurement. Subsequently, the glucose uptake of MCF-7 spheroids was evaluated using living spheroids, fixed spheroids, supernatants, and glucose transporter inhibitor-treated spheroids. Conventionally, the direct optical measurement of glucose uptake requires fluorescence-labeled glucose derivatives. In addition, the glucose uptake can be evaluated by measuring the glucose concentration in the medium by optical or electrochemical measurements. However, glucose needs to be consumed in the entire cell culture medium, which needs a long culture time. In contrast, our system can measure glucose in approximately 5 min without any labels because of in situ electrochemical measurements. This system can be used for in situ measurements in in vitro cell culture systems, including organ-on-a-chip for drug screening.

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Section: ■ Results and Discussionmentioning

confidence: 99%

“…The porous membranes allowed perfusion culture in these experiments. The vertical-flow perfusion through the pores improved cell functions because the flow provided oxygen and nutrients to the spheroids . The porous membrane electrodes can evaluate cell functions while maintaining the cell culture environment.…”

Section: Resultsmentioning

confidence: 99%

Enzyme-Free In-Situ Electrochemical Measurement Using a Porous Membrane Electrode for Glucose Transport into Cell Spheroids

Utagawa,

Ino,

Shinoda

et al. 2024

ACS Sens.

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“…For instance, PDMS-based porous microneedles as a biosensor for continuous monitoring have been proposed. 45 Cell culture devices integrated with porous PDMS sponges have also been reported, 46 in which the direct medium supply to the cells improved cell functions and viability. Fabrication of such 3D structures using polydimethylsiloxane (PDMS) as the material would facilitate the integration and incorporation of the structures into PDMS-based microfluidic devices.…”

Section: Introductionmentioning

confidence: 99%

“…Fabrication of such 3D structures using polydimethylsiloxane (PDMS) as the material would facilitate the integration and incorporation of the structures into PDMS-based microfluidic devices. In fabricating these porous substrates, various types of porogens, including solid particles, [45][46][47][48] water bubbles 49 and ice particles, 50 have been utilized as sacrificial sources for the creation of pores. Among them, the use of uniformly sized spherical particles as the porogen facilitates the formation of so-called inverse colloidal crystals (ICC) or inverse opals, in which the surface/internal pores are uniformly and densely arranged.…”

Section: Introductionmentioning

confidence: 99%

Pushing the limits of microfluidic droplet production efficiency: engineering microchannels with seamlessly implemented 3D inverse colloidal crystals

Mashiyama,

Hemmi,

Sato

et al. 2024

Lab Chip

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“…Specifically, here we focused on porous substrates with interconnected micropores . Recently, studies have reported the fabrication of sponge-like matrices using PDMS as the material and the incorporation of porous structures into microfluidic devices. , We anticipated that a porous substrate with high-density micropores on the surface, once integrated into microchannel structures, could function as a highly efficient drainage system to extract the flow from a particle/cell suspension. From these viewpoints, we developed a new concept of a sheath flow generator, as shown in Figure .…”

Section: Introductionmentioning

confidence: 99%

High-Density Microporous Drainage-Integrating Sheath Flow Generator for Streamlining Microfluidic Cell Sorting Systems

Hayashi,

Hemmi,

Saito

et al. 2024

Anal. Chem.

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Tremendous efforts have been made to develop practical and efficient microfluidic cell and particle sorting systems; however, there are technological limitations in terms of system complexity and low operability. Here, we propose a sheath flow generator that can dramatically simplify operational procedures and enhance the usability of microfluidic cell sorters. The device utilizes an embedded polydimethylsiloxane (PDMS) sponge with interconnected micropores, which is in direct contact with microchannels and seamlessly integrated into the microfluidic platform. The high-density micropores on the sponge surface facilitated fluid drainage, and the drained fluid was used as the sheath flow for downstream cell sorting processes. To fabricate the integrated device, a new process for sponge-embedded substrates was developed through the accumulation, incorporation, and dissolution of PMMA microparticles as sacrificial porogens. The effects of the microchannel geometry and flow velocity on the sheath flow generation were investigated. Furthermore, an asymmetric lattice-shaped microchannel network for cell/particle sorting was connected to the sheath flow generator in series, and the sorting performances of model particles, blood cells, and spiked tumor cells were investigated. The sheath flow generation technique developed in this study is expected to streamline conventional microfluidic cell-sorting systems as it dramatically improves versatility and operability.

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A multiscale, vertical-flow perfusion system with integrated porous microchambers for upgrading multicellular spheroid culture

Abstract: Spheroid formation assisted by microengineered chambers is a versatile approach for morphology-controlled three-dimensional (3D) cell cultivation with physiological relevance to human tissues. However, the limitation in diffusion-based oxygen/nutrition transport has...

Cited by 6 publications

References 65 publications

Enzyme-Free In-Situ Electrochemical Measurement Using a Porous Membrane Electrode for Glucose Transport into Cell Spheroids

Enzyme-Free In-Situ Electrochemical Measurement Using a Porous Membrane Electrode for Glucose Transport into Cell Spheroids

Pushing the limits of microfluidic droplet production efficiency: engineering microchannels with seamlessly implemented 3D inverse colloidal crystals

High-Density Microporous Drainage-Integrating Sheath Flow Generator for Streamlining Microfluidic Cell Sorting Systems

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