Use of Flexible Materials with a Novel Pressure Driven Equibiaxial Cell Stretching Device for Mechanical Stimulation of Single Mammalian Cells

2003 ◽  
Vol 773 ◽  
Author(s):  
James D. Kubicek ◽  
Stephanie Brelsford ◽  
Philip R. LeDuc
Keyword(s):  
Cell Fate ◽  
Mechanical Stimulation ◽  
Mammalian Cells ◽  
Cellular Response ◽  
Single Cells ◽  
Complex Nature ◽  
Cellular Behavior ◽  
Cell Stretching ◽  
Stimulation Of ◽  
Pressure Driven

AbstractMechanical stimulation of single cells has been shown to affect cellular behavior from the molecular scale to ultimate cell fate including apoptosis and proliferation. In this, the ability to control the spatiotemporal application of force on cells through their extracellular matrix connections is critical to understand the cellular response of mechanotransduction. Here, we develop and utilize a novel pressure-driven equibiaxial cell stretching device (PECS) combined with an elastomeric material to control specifically the mechanical stimulation on single cells. Cells were cultured on silicone membranes coated with molecular matrices and then a uniform pressure was introduced to the opposite surface of the membrane to stretch single cells equibiaxially. This allowed us to apply mechanical deformation to investigate the complex nature of cell shape and structure. These results will enhance our knowledge of cellular and molecular function as well as provide insights into fields including biomechanics, tissue engineering, and drug discovery.

scholarly journals Localized mechanical stimulation of single cells with engineered spatio-temporal profile

Lab on a Chip ◽  
2018 ◽  
Vol 18 (19) ◽  
pp. 2955-2965 ◽  
Author(s):  
M. Monticelli ◽  
D. S. Jokhun ◽  
D. Petti ◽  
G. V. Shivashankar ◽  
R. Bertacco
Keyword(s):  
Cell Culture ◽  
Mechanical Stimulation ◽  
Single Cells ◽  
Mechanical Stimuli ◽  
Temporal Profile ◽  
A Cell ◽  
Spatio Temporal ◽  
Stimulation Of

We introduce a new platform for mechanobiology based on active substrates, made of Fe-coated polymeric micropillars, capable to apply mechanical stimuli with tunable spatio-temporal profile on a cell culture.

A novel MEMS device for the multidirectional mechanical stimulation of single cells: Preliminary results

2014 ◽  
Vol 78 ◽  
pp. 131-140 ◽  
Author(s):  
Francesca Antoniolli ◽  
Stefano Maggiolino ◽  
Nicola Scuor ◽  
Paolo Gallina ◽  
Orfeo Sbaizero
Keyword(s):  
Mechanical Stimulation ◽  
Single Cells ◽  
Preliminary Results ◽  
Mems Device ◽  
Stimulation Of

scholarly journals Multiplexed biochemical imaging reveals caspase activation patterns underlying single cell fate

2018 ◽  
Author(s):  
Maximilian W. Fries ◽  
Kalina T. Haas ◽  
Suzan Ber ◽  
John Saganty ◽  
Emma K. Richardson ◽  
...  
Keyword(s):  
Cell Death ◽  
Single Cell ◽  
Cell Fate ◽  
Genetic Heterogeneity ◽  
Cellular Response ◽  
Single Cells ◽  
Caspase Activation ◽  
Cell Fate Decisions ◽  
Network Activation ◽  
Biochemical Imaging

The biochemical activities underlying cell-fate decisions vary profoundly even in genetically identical cells. But such non-genetic heterogeneity remains refractory to current imaging methods, because their capacity to monitor multiple biochemical activities in single living cells over time remains limited1. Here, we deploy a family of newly designed GFP-like sensors (NyxBits) with fast photon-counting electronics and bespoke analytics (NyxSense) in multiplexed biochemical imaging, to define a network determining the fate of single cells exposed to the DNA-damaging drug cisplatin. By simultaneously imaging a tri-nodal network comprising the cell-death proteases Caspase-2, -3 and -92, we reveal unrecognized single-cell heterogeneities in the dynamics and amplitude of caspase activation that signify survival versus cell death via necrosis or apoptosis. Non-genetic heterogeneity in the pattern of caspase activation recapitulates traits of therapy resistance previously ascribed solely to genetic causes3,4. Chemical inhibitors that alter these patterns can modulate in a predictable manner the phenotypic landscape of the cellular response to cisplatin. Thus, multiplexed biochemical imaging reveals cellular populations and biochemical states, invisible to other methods, underlying therapeutic responses to an anticancer drug. Our work develops widely applicable tools to monitor the dynamic activation of biochemical networks at single-cell resolution. It highlights the necessity to resolve patterns of network activation in single cells, rather than the average state of individual nodes, to define, and potentially control, mechanisms underlying cellular decisions in health and disease.

scholarly journals Heterogeneity of Sonic Hedgehog Response Dynamics and Fate Specification in Single Neural Progenitors

2018 ◽  
Author(s):  
Fengzhu Xiong ◽  
Andrea R. Tentner ◽  
Tom W. Hiscock ◽  
Peng Huang ◽  
Sean G. Megason
Keyword(s):  
Cell Fate ◽  
Sonic Hedgehog ◽  
Cellular Response ◽  
Single Cells ◽  
Neural Progenitors ◽  
Significant Heterogeneity ◽  
Response Dynamics ◽  
Output Analysis ◽  
Response Level ◽  
Fate Specification

SUMMARYDuring neural tube patterning, a gradient of Sonic hedgehog (Shh) signaling specifies ventral progenitor fates. The cellular response to Shh is processed through a genetic regulatory network (GRN) to code distinct fate decisions. This process integrates Shh response level, duration and other inputs and is affected by noise in signaling and cell position. How reliably a single cell’s Shh response profile predicts its fate choice is unclear. Here we use live imaging to track neural progenitors that carry both Shh and fate reporters in zebrafish embryos. We found that there is significant heterogeneity between Shh response and fate choice in single cells. We quantitatively modeled reporter intensities to obtain single cell response levels over time and systematically determined their correlation with multiple models of cell fate specification. Our input-output analysis shows that while no single metric perfectly predicts fate choices, the maximal Shh response level correlates best overall with progenitor fate choices across the anterior-posterior axis.

scholarly journals Magneto-active substrates for local mechanical stimulation of living cells

2017 ◽  
Author(s):  
Cécile M. Bidan ◽  
Mario Fratzl ◽  
Alexis Coullomb ◽  
Philippe Moreau ◽  
Alain H. Lombard ◽  
...  
Keyword(s):  
Mechanical Stimulation ◽  
Cellular Response ◽  
Single Cells ◽  
Magnetic Actuation ◽  
Cellular Functions ◽  
Mechanical Responses ◽  
Mechanical Constraints ◽  
Continuous Surface ◽  
And Migration ◽  
Micro Pillars

AbstractCells are able to sense and react to their physical environment by translating a mechanical cue into an intracellular biochemical signal that triggers biological and mechanical responses. This process, called mechanotransduction, controls essential cellular functions such as proliferation and migration. The cellular response to an external mechanical stimulation has been investigated with various static and dynamic systems, so far limited to global deformations or to local stimulation through discrete substrates. To apply local and dynamic mechanical constraints at the single cell scale through a continuous surface, we have developed and modelled magneto-active substrates made of magnetic micro-pillars embedded in an elastomer. Constrained and unconstrained substrates are analysed to map surface stress resulting from the magnetic actuation of the micro-pillars and the adherent cells. These substrates have a rigidity in the range of cell matrices, and the magnetic micro-pillars generate local forces in the range of cellular forces, both in traction and compression. As an application, we followed the protrusive activity of cells subjected to dynamic stimulations. Our magneto-active substrates thus represent a new tool to study mechanotransduction in single cells, and complement existing techniques by exerting a local and dynamic stimulation, traction and compression, through a continuous soft substrate.

scholarly journals Notch1 regulated autophagy controls survival and suppressor activity of activated murine T-regulatory cells

eLife ◽  
2016 ◽  
Vol 5 ◽  
Author(s):  
Nimi Marcel ◽  
Apurva Sarin
Keyword(s):  
Cell Fate ◽  
Nuclear Export ◽  
Mammalian Cells ◽  
Cellular Response ◽  
T Regulatory Cells ◽  
Nuclear Export Signal ◽  
Ectopic Expression ◽  
Notch Pathway ◽  
Regulatory Cells ◽  
Cell Fate Decisions

Cell survival is one of several processes regulated by the Notch pathway in mammalian cells. Here we report functional outcomes of non-nuclear Notch signaling to activate autophagy, a conserved cellular response to nutrient stress, regulating survival in murine natural T-regulatory cells (Tregs), an immune subset controlling tolerance and inflammation. Induction of autophagy required ligand-dependent, Notch intracellular domain (NIC) activity, which controlled mitochondrial organization and survival of activated Tregs. Consistently, NIC immune-precipitated Beclin and Atg14, constituents of the autophagy initiation complex. Further, ectopic expression of an effector of autophagy (Atg3) or recombinant NIC tagged to a nuclear export signal (NIC-NES), restored autophagy and suppressor function in Notch1-/- Tregs. Furthermore, Notch1 deficiency in the Treg lineage resulted in immune hyperactivity, implicating Notch activity in Treg homeostasis. Notch1 integration with autophagy, revealed in these experiments, holds implications for Notch regulated cell-fate decisions governing differentiation.

Mechanical stimulation of human BON cells: Gαq involvement in 5-HT release

2001 ◽  
Vol 120 (5) ◽  
pp. A83-A83
Author(s):  
M KIM ◽  
N JAVED ◽  
F CHRISTOFI ◽  
H COOKE
Keyword(s):  
Mechanical Stimulation ◽  
Bon Cells ◽  
Stimulation Of

A Photoactivatable α5β1-Specific Integrin Ligand

2018 ◽  
Author(s):  
Roshna Vakkeel ◽  
Aleeza Farrukh ◽  
Aranzazu del Campo
Keyword(s):  
Cellular Response ◽  
Light Exposure ◽  
Matrix Composition ◽  
Cellular Behavior ◽  
Dynamic Variations ◽  
In Situ Activation ◽  
Controlled Exposure ◽  
Cellular Behaviour ◽  
Integrin Ligand

In order to study how dynamic changes of α5β1 integrin engagement affect cellular behaviour, photoactivatable derivatives of α5β1 specific ligands are presented in this article. The presence of the photoremovable protecting group (PRPG) introduced at a relevant position for integrin recognition, temporally inhibits ligand bioactivity. Light exposure at cell-compatible dose efficiently cleaves the PRPG and restores functionality. Selective cell response (attachment, spreading, migration) to the activated ligand on the surface is achieved upon controlled exposure. Spatial and temporal control of the cellular response is demonstrated, including the possibility to in situ activation. Photoactivatable integrin-selective ligands in model microenvironments will allow the study of cellular behavior in response to changes in the activation of individual integrins as consequence of dynamic variations of matrix composition.

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