Polymer film-based microwell array platform for long-term culture and research of human bronchial organoids

Baptista, Danielle; Birgani, Zeinab Tahmasebi; Widowski, Helene; Passanha, Fiona; Stylianidis, Vasili; Knoops, Kèvin; Gubbins, Eva; Iriondo, Cinta; Skarp, Kari-Pekka; Rottier, Robbert J.; Wolfs, Tim G. A. M.; Blitterswijk, Clemens van; LaPointe, Vanessa; Habibović, Pamela; Reynaert, Niki L.; Giselbrecht, Stefan; Truckenmüller, Roman

doi:10.1016/j.mtbio.2023.100603

Cited by 3 publications

(1 citation statement)

References 41 publications

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“…To allow the aggregation and maintenance of the spheroids over time, their culture was performed in 3D microwells. The use of thermoformed microwell arrays has been extensively reported to maintain spheroids, organoids, and other 3D cellular constructs , in culture. Advantages of thermoformed microwell chips are the possibility to upscale fabrication and analysis in a high-throughput manner and apply patterned physical, chemical and biological surface or bulk modification .…”

Section: Resultsmentioning

confidence: 99%

Nanofunctionalized Microparticles for Glucose Delivery in Three-Dimensional Cell Assemblies

Fois,

Zengin,

Song

et al. 2024

ACS Appl. Mater. Interfaces

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cell assemblies, such as multicellular spheroids, can be powerful biological tools to closely mimic the complexity of cell−cell and cell−matrix interactions in a native-like microenvironment. However, potential applications of large spheroids are limited by the insufficient diffusion of oxygen and nutrients through the spheroids and, thus, result in the formation of a necrotic core. To overcome this drawback, we present a new strategy based on nanoparticle-coated microparticles. In this study, microparticles function as synthetic centers to regulate the diffusion of small molecules, such as oxygen and nutrients, within human mesenchymal stem cell (hMSC) spheroids. The nanoparticle coating on the microparticle surface acts as a nutrient reservoir to release glucose locally within the spheroids. We first coated the surface of the poly(lactic-co-glycolic acid) (PLGA) microparticles with mesoporous silica nanoparticles (MSNs) based on electrostatic interactions and then formed cell-nanofunctionalized microparticle spheroids. Next, we investigated the stability of the MSN coating on the microparticles' surface during 14 days of incubation in cell culture medium at 37 °C. Then, we evaluated the influence of MSN-coated PLGA microparticles on spheroid aggregation and cell viability. Our results showed the formation of homogeneous spheroids with good cell viability. As a proof of concept, fluorescently labeled glucose (2-NBD glucose) was loaded into the MSNs at different concentrations, and the release behavior was monitored. For cell culture studies, glucose was loaded into the MSNs coated onto the PLGA microparticles to sustain local nutrient release within the hMSC spheroids. In vitro results demonstrated that the local delivery of glucose from MSNs enhanced the cell viability in spheroids during a short-term hypoxic culture. Taken together, the newly developed nanofunctionalized microparticle-based delivery system may offer a versatile platform for local delivery of small molecules within 3D cellular assemblies and, thus, improve cell viability in spheroids.

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Section: Resultsmentioning

confidence: 99%

Nanofunctionalized Microparticles for Glucose Delivery in Three-Dimensional Cell Assemblies

Fois,

Zengin,

Song

et al. 2024

ACS Appl. Mater. Interfaces

Self Cite

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Polymer Design of Microwell Hydrogels Influences Epithelial–Mesenchymal Interactions During Human Bronchosphere Formation

Eiken,

Levine,

Lee

et al. 2024

Advanced NanoBiomed Research

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Bronchospheres have emerged as a promising in vitro model toward probing questions on organ development and disease. Several organoid models, including from airway (e.g., bronchial, tracheal) cells, require three‐dimensional (3D) Matrigel, a complex mouse tumor‐derived matrix that typically leads to heterogeneous size and structures. Synthetic and naturally derived polymeric hydrogels show increased opportunities as an alternative to Matrigel culture. In addition, recent advances in hydrogel‐based microcavities (i.e., microwells) have shown improved control over organoid size, structure, and composition. Here, we build upon this approach and describe the fabrication and characterization of microwell hydrogels based on other polymers, including diacrylated poly(ethylene glycol), agarose, methacrylated gelatin, and norbornene‐modified hyaluronic acid. Using these microwell hydrogels, human bronchial epithelial cells and lung fibroblasts readily assemble into viable cyst‐like bronchospheres. The study shows that the cellular composition regulates the formation and structure of the bronchosphere which also depends on the type and adhesiveness of the hydrogel. Furthermore, both hydrogel type and cellular composition influence the amount and composition of deposited extracellular matrix within the microwells. This hydrogel fabrication platform provides an accessible in vitro culture platform for the formation and growth of bronchospheres which can be extended to the culture of other organoid systems.

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Organoids: Principle, application and perspective

Liu,

Fang,

Aazmi

et al. 2024

TIL

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<p>Organoid technology, a notable advancement in biomedical engineering, has emerged over the past decade, offering significant scientific and therapeutic potential. By accurately mimicking the structural and functional intricacies of human organs at a small scale, organoids have become a groundbreaking tool for exploring basic biological principles, understanding disease mechanisms, and progressing regenerative medicine. Despite the large number of relevant reports, a comprehensive summary of current organoid research updates is needed urgently for interdisciplinary researchers with an interest in constructing biomimetic tissue models. This review presents a thorough look at the diverse fields of organoid research, covering the fundamental principles guiding organoid formation, the broad applications in various biological systems, and the encouraging prospects they bring for future scientific and clinical pursuits.</p>

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Polymer film-based microwell array platform for long-term culture and research of human bronchial organoids

Cited by 3 publications

References 41 publications

Nanofunctionalized Microparticles for Glucose Delivery in Three-Dimensional Cell Assemblies

Nanofunctionalized Microparticles for Glucose Delivery in Three-Dimensional Cell Assemblies

Polymer Design of Microwell Hydrogels Influences Epithelial–Mesenchymal Interactions During Human Bronchosphere Formation

Organoids: Principle, application and perspective

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