3D Printed Multiplexed Competitive Migration Assays with Spatially Programmable Release Sources

Haring, Alexander P.; Thompson, Earl A.; Hernandez, Raymundo; Laheri, Sahil; Harrigan, Megan E.; Lear, Taylor; Sontheimer, Harald; Johnson, Blake N.

doi:10.1002/adbi.201900225

Cited by 4 publications

(3 citation statements)

References 61 publications

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“…To establish flow-free conditions, microfluidic devices can incorporate porous materials such as hydrogels, Matrigel or agarose [24][25][26][27][28][29]. Although, these systems can create stable gradients, their applications have been limited since they often require extended times to establish a gradient due to the relatively low diffusivity in porous materials [30].…”

Section: Introductionmentioning

confidence: 99%

Chemotactic Responses of Jurkat Cells in Microfluidic Flow-Free Gradient Chambers

et al. 2020

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Gradients of soluble molecules coordinate cellular communication in a diverse range of multicellular systems. Chemokine-driven chemotaxis is a key orchestrator of cell movement during organ development, immune response and cancer progression. Chemotaxis assays capable of examining cell responses to different chemokines in the context of various extracellular matrices will be crucial to characterize directed cell motion in conditions which mimic whole tissue conditions. Here, a microfluidic device which can generate different chemokine patterns in flow-free gradient chambers while controlling surface extracellular matrix (ECM) to study chemotaxis either at the population level or at the single cell level with high resolution imaging is presented. The device is produced by combining additive manufacturing (AM) and soft lithography. Generation of concentration gradients in the device were simulated and experimentally validated. Then, stable gradients were applied to modulate chemotaxis and chemokinetic response of Jurkat cells as a model for T lymphocyte motility. Live imaging of the gradient chambers allowed to track and quantify Jurkat cell migration patterns. Using this system, it has been found that the strength of the chemotactic response of Jurkat cells to CXCL12 gradient was reduced by increasing surface fibronectin in a dose-dependent manner. The chemotaxis of the Jurkat cells was also found to be governed not only by the CXCL12 gradient but also by the average CXCL12 concentration. Distinct migratory behaviors in response to chemokine gradients in different contexts may be physiologically relevant for shaping the host immune response and may serve to optimize the targeting and accumulation of immune cells to the inflammation site. Our approach demonstrates the feasibility of using a flow-free gradient chamber for evaluating cross-regulation of cell motility by multiple factors in different biologic processes.

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Section: Introductionmentioning

confidence: 99%

Chemotactic Responses of Jurkat Cells in Microfluidic Flow-Free Gradient Chambers

et al. 2020

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“…Separately, the lack of an optimal concentration in the response of V. alginolyticus to leucine likely indicates a higher saturation threshold in the relevant chemoreceptors, relative to its serine response. This demonstrated screening efficiency highlights the benefits of this new microfluidic platform for tackling large-scale chemotaxis studies in a complementary manner to existing tools ( Lambert et al, 2017 ; Raina et al, 2022 ; Haring et al, 2020 ; Satti et al, 2020 ), for a diverse array of micro-organisms and compounds.…”

Section: Resultsmentioning

confidence: 89%

Multiplexed microfluidic screening of bacterial chemotaxis

Stehnach

Henshaw

Floge

et al. 2023

eLife

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Microorganism sensing of and responding to ambient chemical gradients regulates a myriad of microbial processes that are fundamental to ecosystem function and human health and disease. The development of efficient, high-throughput screening tools for microbial chemotaxis is essential to disentangling the roles of diverse chemical compounds and concentrations that control cell nutrient uptake, chemorepulsion from toxins, and microbial pathogenesis. Here, we present a novel microfluidic multiplexed chemotaxis device (MCD) which uses serial dilution to simultaneously perform six parallel bacterial chemotaxis assays that span five orders of magnitude in chemostimulant concentration on a single chip. We first validated the dilution and gradient generation performance of the MCD, and then compared the measured chemotactic response of an established bacterial chemotaxis system (Vibrio alginolyticus) to a standard microfluidic assay. Next, the MCD’s versatility was assessed by quantifying the chemotactic responses of different bacteria (Psuedoalteromonas haloplanktis, Escherichia coli) to different chemoattractants and chemorepellents. The MCD vastly accelerates the chemotactic screening process, which is critical to deciphering the complex sea of chemical stimuli underlying microbial responses.

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“…While many bioprinted constructs are composed of a single cell type, applications of 3D bioprinting to fabrication of organ-on-a-chip system requires bioprinting of multiple cell types on a single substrate [32][33][34]. In addition to the fabrication of organ-chip platforms, applications of 3D bioprinting to the design of chemotactic signals [35] and production of cell-laden scaffolds for repair of complex injuries [34] difference associated with sensing of bioink cell viability (12 kHz) (see figure 5(a)). While the data in figure 4(a) suggest that one might expect the PC-12 cell-laden bioinks to exhibit a lower impedance than those that contain 3T3 cells based on their relative viabilities, the data in figure 5(a) suggest that other differences among the cell types, such as size and morphology, dominate viability effects in terms of the impedance response.…”

Section: Detection Of Cell Type Differences Among Extruded Cell-laden...mentioning

confidence: 99%

3D bioprinting using hollow multifunctional fiber impedimetric sensors

et al. 2020

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3D bioprinting is an emerging biofabrication process for the production of adherent cell-based products, including engineered tissues and foods. While process innovations are rapidly occurring in the area of process monitoring, which can improve fundamental understanding of process-structure-property relations as well as product quality by closed-loop control techniques, in-line sensing of the bioink composition remains a challenge. Here, we report that hollow multifunctional fibers enable in-line impedimetric sensing of bioink composition and exhibit selectivity for real-time classification of cell type, viability, and state of differentiation during bioprinting. Continuous monitoring of the fiber impedance magnitude and phase angle response from 102 to 106 Hz during microextrusion 3D bioprinting enabled compositional and quality analysis of alginate bioinks that contained fibroblasts, neurons, or mouse embryonic stem cells (mESCs). Fiber impedimetric responses associated with the bioinks that contained differentiated mESCs were consistent with differentiation marker expression characterized by immunocytochemistry. 3D bioprinting through hollow multifunctional fiber impedimetric sensors enabled classification of stem cells as stable or randomly differentiated populations. This work reports an advance in monitoring 3D bioprinting processes in terms of in-line sensor-based bioink compositional analysis using fiber technology and provides a non-invasive sensing platform for achieving future quality-controlled bioprinted tissues and injectable stem-cell therapies.

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3D Printed Multiplexed Competitive Migration Assays with Spatially Programmable Release Sources

Cited by 4 publications

References 61 publications

Chemotactic Responses of Jurkat Cells in Microfluidic Flow-Free Gradient Chambers

Chemotactic Responses of Jurkat Cells in Microfluidic Flow-Free Gradient Chambers

Multiplexed microfluidic screening of bacterial chemotaxis

3D bioprinting using hollow multifunctional fiber impedimetric sensors

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