Microchannel device for proteomic analysis of migrating cancer cells

Bui, Loan; Shen, Qionghua; Hill, Tamara; Bhuiyan, Sayem H; Barakat, Rami; Saavedra, V Oscar; Kong, Calvin; Battiste, James; Kim, Young-tae

doi:10.1088/2057-1976/aacd5f

Cited by 7 publications

(8 citation statements)

References 35 publications

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“…Cytochalasin D, as an actin polymerization disturber, is believed to damage actin by binding to F-actin polymer and preventing the polymerization of actin monomers [10]. From our previous study, actin activity was associated with cancer cell migration and this activity was significantly increased in migrating cancer cell groups [22]. The decreasing velocity associating with increasing the CytoD concentration proved our prior observations and proved the accuracy of our screening system in an mHTS manner.…”

Section: Migrating Inhibition Related Drug Screening and Different Se...supporting

confidence: 77%

Single-cell-level screening method for migratory cancer cells and its potential feasibility in high-throughput manner

Shen¹,

Niederstrasser²,

Barakat³

et al. 2020

Biofabrication

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High-throughput screening (HTS) is a well-established approach for tumor-specific drug development because of its high efficiency and customizable selection of antineoplastic drugs. However, there is still a lack of an appropriate cell-based HTS specific for migratory cancer cells. In the study presented here, we created a novel assay (mHTS): a single-cell-level screening method targeting migratory cancer cells and can be applied in a high-throughput manner. This mHTS platform is based on microchannel devices (providing physical confinement during cell migration and limit migrating cells’ proliferation rate) assembled 96-well plate (fitting to HTS manner). To determine the feasibility of this assay, we quantified the anti-migratory and anti-viability effects of several molecules (Cytochalasin D, Doxorubicin and AZD-6244) on migrating (creeping inside microchannel) glioblastoma multiforme (GBM) cells. After analyzing migration screening data that was collected on a single-cell-level, we were able to compare those drug’s effects on cancer cells’ migration velocity and uncovered the migration inhibiting potential of AZD (500 nM and 1000 nM). Viability data based on single-cell-level screening also allowed us to further understand the same drug’s different lethality toward migrating and normal 2D cultured cancer cells. The Pre-classification of subpopulations enables us to study the heterogeneity of cancer and ensures our method’s feasibility for a high-throughput manner. All these results proved our mHTS platform is suitable for single-cell-level anti-migration drug screening and has potential feasibility in promoting the development of anti-migratory-cancer-drug in a high-throughput manner.

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Section: Migrating Inhibition Related Drug Screening and Different Se...supporting

confidence: 77%

Single-cell-level screening method for migratory cancer cells and its potential feasibility in high-throughput manner

Shen¹,

Niederstrasser²,

Barakat³

et al. 2020

Biofabrication

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“…Microfluidic devices ( Figures 3B,C ) were fabricated using previously established methods ( Bui et al, 2018 ) and were taped to prevent entry of dust into the channels during storage. Prior to use, the devices and coverslips were sterilized in 70% (v/v) ethanol for 15 min, followed by 3 additional washes with sterile double-deionized water inside a biosafety cabinet (Nuaire LabGard ES Class II, Type A2).…”

Section: Methodsmentioning

confidence: 99%

The Opposing Role of Propionate in Modulating Listeria monocytogenes Intracellular Infections

et al. 2021

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Listeria monocytogenes is a Gram-positive, intracellular pathogen responsible for the highly fatal foodborne illness listeriosis. Establishing intracellular infections requires the coordinated expressions of a variety of virulence factors, such as the pore-forming toxin listeriolysin O (LLO), in response to various intra- and extracellular signals. For example, we previously reported that L. monocytogenes differentially modulated LLO production in response to exogenous propionate, a short chain fatty acid either used in salt form as a human food ingredient or produced endogenously by gut microbial fermentation. Therefore, propionate is likely a continuously present signal throughout the L. monocytogenes transmission and infection process. However, little is known about the role of propionate in modulating L. monocytogenes-host interactions. Here we investigated the impact of propionate treatment on L. monocytogenes intracellular infections using cell culture infection models. Propionate treatment was performed separately on L. monocytogenes or host cells before or during infections to better distinguish pathogen-versus-host responses to propionate. Intracellular CFU in RAW264.7 macrophages and plaque diameters in L-fibroblasts were measured as proxy for intracellular infection outcomes. Nitrite levels and cellular morphology were also measured to assess host responses to propionate. We found that propionate pretreatment of anaerobic, but not aerobic, L. monocytogenes significantly enhanced subsequent intracellular infections in both cell types and nitrite production by infected macrophages. Propionate treatment of uninfected macrophages significantly altered cell morphology, seen by longer cells and greater migration, and reduced nitrite concentration in activated macrophages. Treatment of macrophages with propionate prior to or during infections significantly inhibited intracellular growth of L. monocytogenes, including those pre-treated with propionate. These results showcased an opposing effect of propionate on L. monocytogenes intracellular infections and strongly support propionate as an important signaling molecule for both the pathogen and the host cell that can potentially alter the outcome of L. monocytogenes-host interactions.

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“…Microfluidic devices are useful tools for modifying the physical properties of the cell culture to observe physical phenomena in cell response like the migration of tumor cells 64,65 . Microfluidics empower researchers to control the exposure and flow of treatments over cell cultures in a way the other 2D methods covered do not.…”

Section: Two‐dimensional High Throughput Cell Culture Systemsmentioning

confidence: 99%

Advances in high throughput cell culture technologies for therapeutic screening and biological discovery applications

Ryoo,

Kimmel,

Rondo

et al. 2023

Bioengineering & Transla Med

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Cellular phenotypes and functional responses are modulated by the signals present in their microenvironment, including extracellular matrix (ECM) proteins, tissue mechanical properties, soluble signals and nutrients, and cell–cell interactions. To better recapitulate and analyze these complex signals within the framework of more physiologically relevant culture models, high throughput culture platforms can be transformative. High throughput methodologies enable scientists to extract increasingly robust and broad datasets from individual experiments, screen large numbers of conditions for potential hits, better qualify and predict responses for preclinical applications, and reduce reliance on animal studies. High throughput cell culture systems require uniformity, assay miniaturization, specific target identification, and process simplification. In this review, we detail the various techniques that researchers have used to face these challenges and explore cellular responses in a high throughput manner. We highlight several common approaches including two‐dimensional multiwell microplates, microarrays, and microfluidic cell culture systems as well as unencapsulated and encapsulated three‐dimensional high throughput cell culture systems, featuring multiwell microplates, micromolds, microwells, microarrays, granular hydrogels, and cell‐encapsulated microgels. We also discuss current applications of these high throughput technologies, namely stem cell sourcing, drug discovery and predictive toxicology, and personalized medicine, along with emerging opportunities and future impact areas.

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Microchannel device for proteomic analysis of migrating cancer cells

Cited by 7 publications

References 35 publications

Single-cell-level screening method for migratory cancer cells and its potential feasibility in high-throughput manner

Single-cell-level screening method for migratory cancer cells and its potential feasibility in high-throughput manner

The Opposing Role of Propionate in Modulating Listeria monocytogenes Intracellular Infections

Advances in high throughput cell culture technologies for therapeutic screening and biological discovery applications

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