Fabrication approaches for high-throughput and biomimetic disease modeling

Grubb, Mackenzie; Caliari, Steven R.

doi:10.1016/j.actbio.2021.03.006

Cited by 7 publications

(7 citation statements)

References 283 publications

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“…A recent review summarizes the implementation of automation in research laboratories related to biomimetic modeling of diseases and thus enabling high-content studies. 15 Notably, manual pipetting can be replaced by automation, with the advantage of saving labor time. Indeed, recent studies showed that manual pipetting accuracy depends on the individual operator and thus decreases data reproducibility.…”

Section: ■ Introductionmentioning

confidence: 99%

“…17 Standardized multiwell plates with for example 96 wells offer a degree of miniaturization, which reduces the amount of precious cells like stem cells and expensive molecules like growth factors and chemokines while still providing sufficient sample volume and thus are adapted for automated biomaterial fabrication. 15 They are commercially available in different versions with different substrates, mostly plastic and glass, and are compatible with common laboratory equipment, including liquid-handling robots, microplate readers, and high-content screening microscopes. This makes them candidates for automation over the whole experimental workflow from fabrication, cell seeding, sample preparation, data acquisition, and post-treatment.…”

Section: ■ Introductionmentioning

confidence: 99%

“…The researcher would formulate a precise question, later recover the desired biomaterial, and present data to the scientific community, while the central robot takes over the experimental design, biomaterial fabrication, cell culture, and data analysis. A recent review summarizes the implementation of automation in research laboratories related to biomimetic modeling of diseases and thus enabling high-content studies . Notably, manual pipetting can be replaced by automation, with the advantage of saving labor time.…”

Section: Introductionmentioning

confidence: 99%

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Automated Fabrication of Streptavidin-Based Self-assembled Materials for High-Content Analysis of Cellular Response to Growth Factors

Sefkow-Werner

Pennec

Machillot

et al. 2022

ACS Appl. Mater. Interfaces

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The automation of liquid-handling routines offers great potential for fast, reproducible, and labor-reduced biomaterial fabrication but also requires the development of special protocols. Competitive systems demand for a high degree in miniaturization and parallelization while maintaining flexibility regarding the experimental design. Today, there are only a few possibilities for automated fabrication of biomaterials inside multiwell plates. We have previously demonstrated that streptavidin-based biomimetic platforms can be employed to study cellular behaviors on biomimetic surfaces. So far, these self-assembled materials were made by stepwise assembly of the components using manual pipetting. In this work, we introduce for the first time a fully automated and adaptable workflow to functionalize glass-bottom multiwell plates with customized biomimetic platforms deposited in single wells using a liquid-handling robot. We then characterize the cell response using automated image acquisition and subsequent analysis. Furthermore, the molecular surface density of the biomimetic platforms was characterized in situ using fluorescence-based image correlation spectroscopy. These measurements were in agreement with standard ex situ spectroscopic ellipsometry measurements. Due to automation, we could do a proof of concept to study the effect of heparan sulfate on the bioactivity of bone morphogenetic proteins on myoblast cells, using four different bone morphogenetic proteins (BMPs) (2, 4, 6, and 7) in parallel, at five increasing concentrations. Using such an automated self-assembly of biomimetic materials, it may be envisioned to further investigate the role of a large variety of extracellular matrix (ECM) components and growth factors on cell signaling.

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Section: ■ Introductionmentioning

confidence: 99%

Section: ■ Introductionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Automated Fabrication of Streptavidin-Based Self-assembled Materials for High-Content Analysis of Cellular Response to Growth Factors

Sefkow-Werner

Pennec

Machillot

et al. 2022

ACS Appl. Mater. Interfaces

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“…Various methods to increase the throughput of hydrogel fabrication have been developed with unique sets of advantages and disadvantages centered around factors such as fabrication time and complexity . Platforms such as hydrogel-patterned microarrays and microwells, gradient hydrogels, and microgel systems offer high-throughput fabrication but often come with their own limitations.…”

Section: Introductionmentioning

confidence: 99%

Modular Multiwell Viscoelastic Hydrogel Platform for Two- and Three-Dimensional Cell Culture Applications

Skelton,

Gentry,

Astrab

et al. 2024

ACS Biomater. Sci. Eng.

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Hydrogels have gained significant popularity as model platforms to study reciprocal interactions between cells and their microenvironment. While hydrogel tools to probe many characteristics of the extracellular space have been developed, fabrication approaches remain challenging and time-consuming, limiting multiplexing or widespread adoption. Thus, we have developed a modular fabrication approach to generate distinct hydrogel microenvironments within the same 96-well plate for increased throughput of fabrication as well as integration with existing high-throughput assay technologies. This approach enables in situ hydrogel mechanical characterization and is used to generate both elastic and viscoelastic hydrogels across a range of stiffnesses. Additionally, this fabrication method enabled a 3-fold reduction in polymer and up to an 8-fold reduction in fabrication time required per hydrogel replicate. The feasibility of this platform for two-dimensional (2D) cell culture applications was demonstrated by measuring both population-level and single-cell-level metrics via microplate reader and high-content imaging. Finally, a 96-well hydrogel array was utilized for three-dimensional (3D) cell culture, demonstrating the ability to support high cell viability. Together, this work demonstrates a versatile and easily adaptable fabrication approach that can support the ever-expanding tool kit of hydrogel technologies for cell culture applications.

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“…Various methods of hydrogel fabrication with increased throughput have been developed with unique sets of advantages and disadvantages centered around factors such as fabrication time and complexity 9 . Platforms such as microarrays and microwells offer adequate throughput required for large scale drug screens, but often with little to no biomimetic complexity.…”

Section: Introductionmentioning

confidence: 99%

Modular multiwell viscoelastic hydrogel platform for two- and three-dimensional cell culture applications

Skelton,

Gentry,

Astrab

et al. 2023

Preprint

Self Cite

View full text Add to dashboard Cite

Hydrogels have gained significant popularity as model platforms to study the reciprocal interactions between cells and their microenvironment. While hydrogel tools to probe many characteristics of the extracellular space have been developed, fabrication approaches remain challenging and time-consuming, limiting multiplexing or widespread adoption. Thus, we have developed a modular fabrication approach to generate distinct hydrogel microenvironments within 96-well plates for increased throughput of fabrication as well as integration with existing high-throughput assay technologies. This approach enablesin situhydrogel mechanical characterization and was used to generate both elastic and viscoelastic hydrogels across a range of stiffnesses. Additionally, this fabrication method enabled a 3-fold reduction in polymer and up to an 8-fold reduction in fabrication time required per hydrogel replicate. The feasibility of this platform for cell culture applications was demonstrated by measuring both population-level and single cell-level metrics via microplate reader and high-content imaging. Finally, the 96-well hydrogel array was utilized for 3D cell culture, demonstrating the ability to support high cell viability. Together, this work demonstrates a versatile and easily adoptable fabrication approach that can support the ever-expanding tool kit of hydrogel technologies for cell culture applications.

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Fabrication approaches for high-throughput and biomimetic disease modeling

Cited by 7 publications

References 283 publications

Automated Fabrication of Streptavidin-Based Self-assembled Materials for High-Content Analysis of Cellular Response to Growth Factors

Automated Fabrication of Streptavidin-Based Self-assembled Materials for High-Content Analysis of Cellular Response to Growth Factors

Modular Multiwell Viscoelastic Hydrogel Platform for Two- and Three-Dimensional Cell Culture Applications

Modular multiwell viscoelastic hydrogel platform for two- and three-dimensional cell culture applications

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