A microfluidic dual gradient generator for conducting cell-based drug combination assays

Kilinc, Devrim; Schwab, Jefrem; Rampini, Stefano; Ikpekha, Oshoke W.; Thampi, Ashwin; Blasiak, Agata; Li, Peng; Schwamborn, Robert; Kölch, Walter; Matallanas, David; G, Lee

doi:10.1039/c5ib00209e

Cited by 27 publications

(23 citation statements)

References 33 publications

(38 reference statements)

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“…We also speculate that this approach will be helpful in understanding several other signaling mechanisms, and it also has potential for applications in pharmacology, and synthetic biology. With the advancements in microfabrication technology 79–83 and mathematical modeling, we speculate that investigating signaling pathways with this approach will provide better understanding of such pathways and also lead to several biomedical applications. Advanced mathematical approaches such as multi-scale modeling and agent-based modeling will further push the investigation at systems scale 84,85 .…”

Section: Discussionmentioning

confidence: 99%

New insights into mammalian signaling pathways using microfluidic pulsatile inputs and mathematical modeling

2017

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Temporally modulated input mimics physiology. This chemical communication strategy filters the biochemical noise through entrainment and phase-locking. Under laboratory conditions, it also expands the observability space for downstream responses. A combined approach involving microfluidic pulsatile stimulation and mathematical modeling has led to deciphering of hidden/unknown temporal motifs in several mammalian signaling pathways and has provided mechanistic insights, including how these motifs combine to form distinct band-pass filters and govern fate regulation under dynamic microenvironment. This approach can be utilized to understand signaling circuit architectures and to gain mechanistic insights for several other signaling systems. Potential applications include synthetic biology and biotechnology, in developing pharmaceutical interventions, and in developing lab-on-chip models.

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Section: Discussionmentioning

confidence: 99%

New insights into mammalian signaling pathways using microfluidic pulsatile inputs and mathematical modeling

2017

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“…One end of the synaptic chamber bifurcates into two branches, one of which terminates in a triangular shape. This terminus is connected to a diamond-shaped co-culture chamber (based on a previous design) ( Kilinc et al, 2016) via 4 μm high, 10 μm wide, and 100 μm long parallel microchannels.…”

Section: Microfluidic Device Design and Fabricationmentioning

confidence: 99%

Pyk2 Overexpression in Postsynaptic Neurons Blocks Aβ_1-42-induced Synaptotoxicity in a Microfluidic Co-Culture Model

Kilinc

Vreulx

Mendes

et al. 2019

Preprint

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Recent meta-analyses of genome-wide association studies identified a number of genetic risk factors ofAlzheimer's disease (AD); however, little is known about the mechanisms by which they contribute to the pathological process. As synapse loss is observed at the earliest stage of AD, deciphering the impact of AD risk genes on synapse formation and maintenance is of great interest. In this paper, we report a microfluidic co-culture device that physically isolates synapses from pre-and postsynaptic neurons and chronically exposes them to toxic amyloid-beta (Aβ) peptides secreted by model cell lines overexpressing wild-type or mutated (V717I) amyloid precursor protein (APP). Co-culture with cells overexpressing mutated APP exposed the synapses of primary hippocampal neurons to Aβ 1-42 molecules at nanomolar concentrations and induced a significant decrease in synaptic connectivity, as evidenced by distance-based assignment of postsynaptic puncta to presynaptic puncta. Treating the cells with antibodies that target different forms of Aβ suggested that low molecular weight oligomers are the likely culprit. As proof of concept, we demonstrate that overexpression of Pyk2 -an AD risk gene involved in synaptic plasticity and shown to decrease in AD brains at gene expression and protein levels-selectively in postsynaptic neurons is protective against Aβ 1-42 -induced synaptotoxicity. In summary, our lab-on-a-chip device provides a physiologically-relevant model of AD-related synaptotoxicity, optimal for assessing the impact of risk genes in pre-and postsynaptic compartments. ExperimentalOligimerization of synthetic Aβ peptides Aβ peptides were oligomerized according to established protocols (4, 32) with minor modifications. The inactive control peptide (Aβ 42-1 ; Abcam; Cambridge, UK) has the same composition as the Aβ 1-42 peptide (California Peptide Research; Napa, CA) but with an inverted amino-acid sequence, and has been widely used as control for oligomeric Aβ 1-42 since it is also prone to oligomerization (33-35). Aβ 1-42 and ICP were treated with hexafluoroisopropanol (Sigma-Aldrich, Saint Louis, MO) to maintain the oligomeric structure and to reduce fibril formation (4), according to manufacturer's instructions. The peptides were resuspended in 1 ml hexafluoroisopropanol and incubated for 1 h at RT, with occasional moderate vortexing, followed by

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“…(D) diffusion‐based microfluidic device with design for bidimensional gradient generation. Kilinc et al (2016) . Reproduced with permission from Ref.…”

Section: Microfluidics: Approaches For Industrial Biotechnologymentioning

confidence: 99%

Microfluidic tools toward industrial biotechnology

Oliveira

Pessoa

Bastos

et al. 2016

Biotechnology Progress

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Microfluidics is a technology that operates with small amounts of fluids and makes possible the investigation of cells, enzymes, and biomolecules and encapsulation of biocatalysts in a greater variety of conditions than permitted using conventional methods. This review discusses technological possibilities that can be applied in the field of industrial biotechnology, presenting the principal definitions and fundamental aspects of microfluidic parameters to better understand advanced approaches. Specifically, concentration gradient generators, droplet-based microfluidics, and microbioreactors are explored as useful tools that can contribute to industrial biotechnology. These tools present potential applications, inclusive as commercial platforms to optimizing in bioprocesses development as screening cells, encapsulating biocatalysts, and determining critical kinetic parameters. © 2016 American Institute of Chemical Engineers Biotechnol. Prog., 32:1372-1389, 2016.

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A microfluidic dual gradient generator for conducting cell-based drug combination assays

Cited by 27 publications

References 33 publications

New insights into mammalian signaling pathways using microfluidic pulsatile inputs and mathematical modeling

New insights into mammalian signaling pathways using microfluidic pulsatile inputs and mathematical modeling

Pyk2 Overexpression in Postsynaptic Neurons Blocks Aβ_1-42-induced Synaptotoxicity in a Microfluidic Co-Culture Model

Microfluidic tools toward industrial biotechnology

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A microfluidic dual gradient generator for conducting cell-based drug combination assays

Cited by 27 publications

References 33 publications

New insights into mammalian signaling pathways using microfluidic pulsatile inputs and mathematical modeling

New insights into mammalian signaling pathways using microfluidic pulsatile inputs and mathematical modeling

Pyk2 Overexpression in Postsynaptic Neurons Blocks Aβ1-42-induced Synaptotoxicity in a Microfluidic Co-Culture Model

Microfluidic tools toward industrial biotechnology

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Product

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Pyk2 Overexpression in Postsynaptic Neurons Blocks Aβ_1-42-induced Synaptotoxicity in a Microfluidic Co-Culture Model