Versatile on-stage microfluidic system for long term cell culture, micromanipulation and time lapse assays

Huang, Yao‐Xiong; He, Can; Wang, Ping; Pan, Yan-Ting; Tuo, Wei-Wei; Yao, Cheng-Can

doi:10.1016/j.bios.2017.09.041

Cited by 5 publications

(4 citation statements)

References 25 publications

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Section: Discussionmentioning

confidence: 99%

“…Previous works on developing the incubatorindependent cell culture system have been mainly designed for continuous microscopy readouts from cultured cells for hours or days [28,30,33,35,43,47,91]. For instance, stage-top mini-incubators have been designed to supply CO 2 , temperature and humidity to the perfusion chambers assembled on the microscope stage [43,45,47,51,53]. To capture neuronal network activity either on-stage incubators are modified to accommodate the MEA amplifiers or custom-developed perfusion chambers are integrated with MEA electrophysiology setup [45,47,[51][52][53].…”

Section: Discussionmentioning

confidence: 99%

“…Different types of on-stage incubators are commercially available that are equipped with self-contained imaging systems to record morphological changes over time or in large sample sizes [42]. Even though, miniaturized on-stage incubators provide an excellent platform for long-term microscopy, these systems still require bulky accessories to provide gas and humidity that makes it difficult to be integrated with screening tools like electrophysiology recording setups [31,43]. Incubator-independent screening platforms aim to provide long-term continuous access to the network microscopy and electrophysiology data, and to exclude disturbances related to the media exchange and handling [31,[44][45][46].…”

Section: Introductionmentioning

confidence: 99%

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Incubator-independent perfusion system integrated with microfluidic device for continuous electrophysiology and microscopy readouts

Habibey¹

2023

Biofabrication

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Advances in primary and stem cell derived neuronal cell culture techniques and abundance of available neuronal cell types have enabled in vitro neuroscience as a substantial approach to model in vivo neuronal networks. Survival of the cultured neurons is inevitably dependent on the cell culture incubators to provide stable temperature and humidity and to supply required CO2 levels for controlling the pH of culture medium. Therefore, imaging and electrophysiology recordings outside of the incubator are often limited to the short-term experimental sessions. This restricts our understanding of physiological events to the short snapshots of recorded data while the major part of temporal data is neglected. Multiple custom-made and commercially available platforms like integrated on-stage incubators have been designed to enable long-term microscopy. Nevertheless, long-term high-spatiotemporal electrophysiology recordings from developing neuronal networks needs to be addressed. In the present work an incubator-independent polydimethylsiloxane (PDMS)-based double-wall perfusion chamber was designed and integrated with multi-electrode arrays (MEAs) electrophysiology and compartmentalized microfluidic device to continuously record from engineered neuronal networks at sub-cellular resolution. Cell culture media underwent iterations of conditioning to the ambient CO2 and adjusting its pH to physiological ranges to retain a stable pH for weeks outside of the incubator. Double-wall perfusion chamber and an integrated air bubble trapper reduced media evaporation and osmolality drifts of the conditioned media for two weeks. Aligned microchannel-microfluidic device on MEA electrodes allowed neurite growth on top of the planar electrodes and amplified their extracellular activity. This enabled continuous sub-cellular resolution imaging and electrophysiology recordings from developing networks and their growing neurites. The on-chip versatile and self-contained system provides long-term, continuous and high spatiotemporal access to the network data and offers a robust in vitro platform with many potentials to be applied on advanced cell culture systems including organ-on-chip and organoid models.

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Section: Discussionmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Incubator-independent perfusion system integrated with microfluidic device for continuous electrophysiology and microscopy readouts

Habibey¹

2023

Biofabrication

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“…The second level is biomimetic microfluidic cell culture, which enables the sterility condition because the closed microfluidic system provides better insulation from the external environment (Huang et al, 2018). Culture systems can better mimic a dynamic environment replicated physiological flow conditions (Zang et al, 2012) to bridge the gap between conventional cultures and complex native in vivo environments (Kieninger et al, 2018).…”

mentioning

confidence: 99%

A novel method of cell culture based on the microfluidic chip for regulation of cell density

Zhang

Wei

et al. 2020

Biotech & Bioengineering

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The regulation of cell density is an important segment in microfluidic cell culture, particularly in the repeated assays. Traditionally, consistent cell density is difficult to achieve, owing to the inaccurate regulation of cell density with manual feedback. A novel cell culture method with automatic feedback is proposed for real‐time regulation of cell density based on microfluidic chip in this paper. Here, an integrated microfluidic system combining cell culture, density detection, and control of proliferation rate was developed. Interdigital electrode structures were sputtered on the microchannel automatically to provide the real‐time feedback information of impedance. The most sensitive frequency was studied to improve the detection resolution of the sensing chip. Cells were cultured on the chip surface and cell density was detected by monitoring the alternation of the impedance. The feedback controller was established by the least squares support vector machines. Then, the cell proliferation rate was automatically controlled using the feedback controller to achieve the desired cell density in the repeated assays. The results show that the standard error of this method is 2.8% indicating that the method can keep a consistency of cell density in the repeated assays. This study provides a basis for improving the accuracy and repeatability in the further assays of finding the optimal drug concentration.

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Tunable and Reversible Gelatin‐Based Bonding for Microfluidic Cell Culture

et al. 2019

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Microfluidic cell culture is widely used to develop biochips and biosensors, culturing and experimenting with cells at the microscale. However, only a very small subset of the existing polymers is currently used in microfluidics. This is mostly due to limitations in reversibility and gas-permeability on the sealant. Hence, the development of a novel bonding technique can enable new applications and uses of plastics in microfluidic cell culture, complementing the omnipresent polydimethylsiloxane (PDMS) for critical applications where harder or non-porous materials are required. The present paper describes a reversible gelatin-based room-temperature method for bonding separate substrates, which enables the sealing of commonly used materials in microfluidics such as thermoplastics, but also elastomers and photopolymers. For most materials, the bonding chip resisted to at least 0.1 MPa. To show the versatility of the described method we bonded microchannels of different sizes, up to 200 μm, and round microstructures.The applicability to cell culture was investigated by culturing colorectal cancer HT-29 cells within the chip. Finally, the cells viability was analyzed by in situ live/dead fluorescence staining. Advantageously, the proposed bonding process is reversible and make possible to tune the permeability of the gelatin layer integrated on chip. This room-temperature bonding method is highly efficient for cell culture in plastic chips, potentially opening new routes for the development of innovative BioMEMS devices.

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Versatile on-stage microfluidic system for long term cell culture, micromanipulation and time lapse assays

Cited by 5 publications

References 25 publications

Incubator-independent perfusion system integrated with microfluidic device for continuous electrophysiology and microscopy readouts

Incubator-independent perfusion system integrated with microfluidic device for continuous electrophysiology and microscopy readouts

A novel method of cell culture based on the microfluidic chip for regulation of cell density

Tunable and Reversible Gelatin‐Based Bonding for Microfluidic Cell Culture

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