Imaging-guided bioreactor for de-epithelialization and long-term cultivation of <i>ex vivo</i> rat trachea

Mir, Seyed Mohammad; Chen, Jiawen; Pinezich, Meghan R.; O’Neill, John D.; Huang, Sarah X.L.; Vunjak-Novakovic, Gordana; Kim, Jinho

doi:10.1039/d1lc01105g

Cited by 6 publications

(6 citation statements)

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Section: Cellular Physiologymentioning

confidence: 99%

“…Unlike traditional visualization techniques involving the optical microscopic method, direct imaging of the culture niche can be a powerful tool for the intrinsic understanding of the cell microenvironment. [18,28,38,39] As one of the examples, in situ, visualization on the organ-onchip designed for airway tissue monitoring was suggested by Mir et al [26] The developed monitoring chip platform comprises the gradient-index (GRIN) lens and imagingenabled PTFE custom valve type bioreactor (Figure 2b). Stem cell differentiation on tissue was visualized by monitoring with bright field and fluorescence imaging inside the tissue that possesses the capability of an imaging-assisted bioreactor platform.…”

Section: Cellular Physiologymentioning

confidence: 99%

Section: Quartz Crystal Microbalancementioning

confidence: 99%

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Advances in sensor developments for cell culture monitoring

Kim,

Yeo

2023

BMEMat

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Cell culture encompasses procedures for extracting cells from their natural tissue and cultivating them under controlled artificial conditions. During this process, various factors, including cell physiological/morphological properties, culture environments, metabolites, and contaminants, have to be precisely controlled and monitored for the survival of cells and the pursuit of the desired properties of the cells. This review summarizes recent advances in sensor technologies and manufacturing strategies for various cell culture platforms using traditional plastics, microfluidic chips, and scalable bioreactors. We share the details of newly developed biological sensors, chemical sensors, optical sensors, electronic chip technologies, and material integration methods. The precise control of parameters based on the feedback by these sensors and electronics enhances cell culture quality and throughput.

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Section: Cellular Physiologymentioning

confidence: 99%

Section: Cellular Physiologymentioning

confidence: 99%

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Advances in sensor developments for cell culture monitoring

Kim,

Yeo

2023

BMEMat

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“…In the current study, we characterized a ferret tracheal explant model of injury and cell engraftment. As opposed to mice, rats, and rabbits, for which tracheal explants were described previously ( 8 – 10 ), ferrets are useful, because they have extensively developed submucosal glands (SMG) throughout cartilaginous airways, which is also the case in humans. This is critical because these structures contribute significantly to airway regeneration ( 11 , 12 ).…”

Section: Introductionmentioning

confidence: 99%

Development and characterization of ferret ex vivo tracheal injury and cell engraftment model

et al. 2023

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The field of airway biology research relies primarily on in vitro and in vivo models of disease and injury. The use of ex vivo models to study airway injury and cell-based therapies remains largely unexplored although such models have the potential to overcome certain limitations of working with live animals and may more closely replicate in vivo processes than in vitro models can. Here, we characterized a ferret ex vivo tracheal injury and cell engraftment model. We describe a protocol for whole-mount staining of cleared tracheal explants, and showed that it provides a more comprehensive structural overview of the surface airway epithelium (SAE) and submucosal glands (SMGs) than 2D sections, revealing previously underappreciated structural anatomy of tracheal innervation and vascularization. Using an ex vivo model of tracheal injury, we evaluated the injury responses in the SAE and SMGs that turned out to be consistent with published in vivo work. We used this model to assess factors that influence engraftment of transgenic cells, providing a system for optimizing cell-based therapies. Finally, we developed a novel 3D-printed reusable culture chamber that enables live imaging of tracheal explants and differentiation of engrafted cells at an air-liquid interface. These approaches promise to be useful for modeling pulmonary diseases and testing therapies.Graphical abstract1,2. We describe here a method for differential mechanical injury of ferret tracheal explants that can be used to evaluate airway injury responses ex vivo. 3. Injured explants can be cultured at ALI (using the novel tissue-transwell device on the right) and submerged long-term to evaluate tissue-autonomous regeneration responses. 4. Tracheal explants can also be used for low throughput screens of compounds to improve cell engraftment efficiency or can be seeded with particular cells to model a disease phenotype. 5. Lastly, we demonstrate that ex vivo-cultured tracheal explants can be evaluated by various molecular assays and by immunofluorescent imaging that can be performed live using our custom-designed tissue-transwell.

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“…Wei et al combined an external lens with a mobile phone into a compact imaging system for imaging and length quantification of single molecule DNA strands [20]. In order to visualize the removal of endogenous epithelium and local delivery of exogenous cells in situ, Kim et al developed a customized micro optical imaging device integrated with a bioreactor [21]. This device has shown great potential in space cell imaging, but the system complexity is high and is not suitable for simple cell culture monitoring.…”

Section: Introductionmentioning

confidence: 99%

A Cell State Monitoring System with Integrated In Situ Imaging and pH Detection

Li,

Zhang,

et al. 2023

Sensors

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Cell models are one of the most widely used basic models in biological research, and a variety of in vitro cell culture techniques and models have been developed recently to simulate the physiological microenvironment in vivo. However, regardless of the technique or model, cell culture is the most fundamental but crucial component. As a result, we have developed a cell culture monitoring system to assess the functional status of cells within a biochip. This article focuses on a mini-microscope made from a readily available camera for in situ continuous observation of cell growth within a biochip and a pH sensor based on optoelectronic sensing for measuring pH. With the aid of this monitoring system, scientists can keep an eye on cell growth in real time and learn how the pH of the culture medium affects it. This study offers a new approach for tracking cells on biochips and serves as a valuable resource for enhancing cell culture conditions.

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Imaging-guided bioreactor for de-epithelialization and long-term cultivation of ex vivo rat trachea

Abstract: Recent synergistic advances in organ-on-chip and tissue engineering technologies offer opportunities to create in vitro-grown tissue or organ constructs that can faithfully recapitulate their in vivo counterparts. Such in vitro...

Cited by 6 publications

References 76 publications

Advances in sensor developments for cell culture monitoring

Advances in sensor developments for cell culture monitoring

Development and characterization of ferret ex vivo tracheal injury and cell engraftment model

A Cell State Monitoring System with Integrated In Situ Imaging and pH Detection

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