Versatile method to generate multiple types of micropatterns

Segerer, Felix J.; Röttgermann, Peter J. F.; Alberola, Alicia Piera; Zahler, Stefan; Rädler, Joachim O.

doi:10.1116/1.4940703

Cited by 22 publications

(15 citation statements)

References 48 publications

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“…In order to simplify the assessment and comparison of motility-related parameters, micropatterning techniques can be exploited to create standardized cell environments and restrict cell migration to defined geometries 35 36 37 . The resulting cellular shapes and dynamics can in turn be compared to theoretical modeling 38 .…”

mentioning

confidence: 99%

Ring-Shaped Microlanes and Chemical Barriers as a Platform for Probing Single-Cell Migration

Schreiber

Segerer

Wagner

et al. 2016

Sci Rep

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Quantification and discrimination of pharmaceutical and disease-related effects on cell migration requires detailed characterization of single-cell motility. In this context, micropatterned substrates that constrain cells within defined geometries facilitate quantitative readout of locomotion. Here, we study quasi-one-dimensional cell migration in ring-shaped microlanes. We observe bimodal behavior in form of alternating states of directional migration (run state) and reorientation (rest state). Both states show exponential lifetime distributions with characteristic persistence times, which, together with the cell velocity in the run state, provide a set of parameters that succinctly describe cell motion. By introducing PEGylated barriers of different widths into the lane, we extend this description by quantifying the effects of abrupt changes in substrate chemistry on migrating cells. The transit probability decreases exponentially as a function of barrier width, thus specifying a characteristic penetration depth of the leading lamellipodia. Applying this fingerprint-like characterization of cell motion, we compare different cell lines, and demonstrate that the cancer drug candidate salinomycin affects transit probability and resting time, but not run time or run velocity. Hence, the presented assay allows to assess multiple migration-related parameters, permits detailed characterization of cell motility, and has potential applications in cell biology and advanced drug screening.Migrating cells play a pivotal role in morphogenesis 1 , immune responses 2 , and cancer metastasis 3 . Their style of motion, often assigned as crawling, is powered by complex cytoskeletal rearrangements that deform and propel the cell. On solid surfaces, eukaryotic cells extend protrusions, which attach to the substrate and are then actively retracted, thus dragging the cell forward. The formation of the leading protrusion of a migrating cell, the lamellipodium, is driven by actin polymerization, while adhesion and contraction are predominantly regulated by integrin-based focal adhesions and the actomyosin apparatus 4,5 . Coupling of focal adhesion complexes to the cytoskeletal network in turn reinforces actin assembly and hence lamellipodia extension 6 . The complex interplay between actomyosin contractility and focal adhesions, which are capable of sensing and transducing chemical and mechanical cues in the extracellular environment, renders the cell sensitive to external stimuli such as the composition and rigidity of the extracellular matrix (ECM) and the underlying substrate 7,8 .In recent studies, various theoretical models for cell migration have been proposed and implemented. These implementations range from molecular level approaches, which describe cell migration in terms of internal reaction diffusion dynamics 9-11 to coarse grained approaches in which individual cells are resembled by sets of pixels 12-14 or interacting, self-propelled geometrical objects [15][16][17] . Many of these models are able to reproduce the basic ...

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mentioning

confidence: 99%

Ring-Shaped Microlanes and Chemical Barriers as a Platform for Probing Single-Cell Migration

Schreiber

Segerer

Wagner

et al. 2016

Sci Rep

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“…Patterning adhesive islands on the surface, as done in 31,46 , would overcome the limitations related to the positioning and orientation of the cells with respect to the pillars. A strategic functionalization of the magneto-active substrate would then guarantee the repeatability of cell experiments by controlling the degree of spreading and the geometry of the cell.…”

Section: Discussionmentioning

confidence: 99%

Magneto-active substrates for local mechanical stimulation of living cells

Bidan

Fratzl

Coullomb

et al. 2017

Preprint

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“…Single-cell microarrays were produced by microscale plasma-induced protein patterning (µPIPP) like already described elsewhere (38,39). The array consists of a micropattern with cell adhesive squares, which are coated with fibronectin.…”

Section: Single-cell Array Fabricationmentioning

confidence: 99%

Correlation of mRNA delivery timing and protein expression in lipid-based transfection

Reiser

Woschée

Mehrotra

et al. 2019

Preprint

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Non-viral gene delivery is constrained by the dwell time that most synthetic nucleic acid nanocarriers spend inside endosomal compartments. In order to overcome this endosomalrelease bottleneck, methods are required that measure nanocarrier uptake kinetics and transfection efficiency simultaneously. Here, we employ live-cell imaging on single-cell arrays (LISCA) to study the delivery-time distribution of lipid-based mRNA complexes under varied serum conditions. By fitting a translation-maturation model to hundreds of individual eGFP reporter fluorescence time courses, the protein expression onset times and the expression rates after transfection are determined. Using this approach, we find that delivery timing and protein expression rates are not intrinsically correlated at the single-cell level, even though population-averaged values of both parameters conjointly change as a function of increasing external serum protein fraction. Lipofectamine mediated delivery showed decreased transfection efficiency and longer delivery times with increasing serum protein concentration. This is in contrast to ionizable lipid nanoparticles (LNPs) mediated transfer, which showed increased efficiency and faster uptake in the presence of serum. In conclusion, the interdependences of single-cell expression rates and onset timing provide additional clues on uptake and release mechanisms, which are useful for improving nucleic acid delivery. author/funder. All rights reserved. No reuse allowed without permission.

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Versatile method to generate multiple types of micropatterns

Cited by 22 publications

References 48 publications

Ring-Shaped Microlanes and Chemical Barriers as a Platform for Probing Single-Cell Migration

Ring-Shaped Microlanes and Chemical Barriers as a Platform for Probing Single-Cell Migration

Magneto-active substrates for local mechanical stimulation of living cells

Correlation of mRNA delivery timing and protein expression in lipid-based transfection

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