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Title Probing cellular mechano-sensitivity using biomembrane-mimicking cell substrates of adjustable stiffness
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Publication Date
Date Available
University/Publisher IUPUI
Abstract

Indiana University-Purdue University Indianapolis (IUPUI)

It is increasingly recognized that mechanical properties of substrates play a pivotal role in the regulation of cellular fate and function. However, the underlying mechanisms of cellular mechanosensing still remain a topic of open debate. Traditionally, advancements in this field have been made using polymeric substrates of adjustable stiffness with immobilized linkers. While such substrates are well suited to examine cell adhesion and migration in an extracellular matrix environment, they are limited in their ability to replicate the rich dynamics found at cell-cell interfaces. To address this challenge, we recently introduced a linker-functionalized polymer-tethered multi-bilayer stack, in which substrate stiffness can be altered by the degree of bilayer stacking, thus allowing the analysis of cellular mechanosensitivity. Here, we apply this novel biomembrane-mimicking cell substrate design to explore the mechanosensitivity of C2C12 myoblasts in the presence of cell-cell-mimicking N-cadherin linkers. Experiments are presented, which demonstrate a relationship between the degree of bilayer stacking and mechanoresponse of plated cells, such as morphology, cytoskeletal organization, cellular traction forces, and migration speed. Furthermore, we illustrate the dynamic assembly of bilayer-bound N-cadherin linkers underneath cellular adherens junctions. In addition, properties of individual and clustered N-cadherins are examined in the polymer-tethered bilayer system in the absence of plated cells. Alternatively, substrate stiffness can be adjusted by the concentration of lipopolymers in a single polymer-tethered lipid bilayer. On the basis of this alternative cell substrate concept, we also discuss recent results on a linker-functionalized single polymer-tethered bilayer substrate with a lateral gradient in lipopolymer concentration (substrate viscoelasticity). Specifically, we show that the lipopolymer gradient has a notable impact on spreading, cytoskeletal organization, and motility of 3T3 fibroblasts. Two cases are discussed: 1. polymer-tethered bilayers with a sharp boundary between low and high lipopolymer concentration regions and 2. polymer-tethered bilayers with a gradual gradient in lipopolymer concentration.

Subjects/Keywords Artificial Substrate; Cadherin; Polymer-tethered Bilayer; Biomembrane-mimicking; Lipid Bilayer; Cadherins  – Research; Cell junctions  – Research; Polymers  – Surfaces; Bilayer lipid membranes  – Research; Cell adhesion  – Research  – Analysis; Biological interfaces  – Research  – Analysis; Cellular control mechanisms  – Research; Cells  – Mechanical properties  – Research; Polymers  – Rheology; Artificial cells  – Research
Contributors Naumann, Christoph A.; Das, Chittaranjan; Thompson, David; Long, Eric C.
Language en
Country of Publication us
Record ID handle:1805/9965
Other Identifiers 10.7912/C22G65
Repository iupui
Date Retrieved
Date Indexed 2020-01-07

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…Extracellular Matrix. EDC/NHS 1-Ethyl-3-(3-dimethylaminopropyl)-carbodiimide /N-hydroxysuccinimide. EPI Epifluorescence. FA Focal adhesion. FAK Focal adhesion kinase. FBS Fetal bovine serum. xv FCS Fluorescence correlation spectroscopy…

…However, the underlying mechanisms of cellular mechanosensing still remain a topic of open debate. Traditionally, advancements in this field have been made using polymeric substrates of adjustable stiffness with immobilized linkers. While such substrates…

…are well suited to examine cell adhesion and migration in an extracellular matrix environment, they are limited in their ability to replicate the rich dynamics found at cell-cell interfaces. To address this challenge, we recently introduced a linker…

…signaling mechanisms that impact cell function and pathology1,2. Meanwhile, it has been recognized that mechanical signals may also have a profound impact on cellular fate and function. Such mechanical cues may include shear stress, osmotic forces…

…mechanoresponse, and in turn different cytoskeleton organization. Research of artificial substrate design and fabrication for the investigation of mechanoresponse between cell forces related from cytoskeleton, such as F-actin, to focal adhesion structure can…

…the linker immobilization may hinder the lateral assembly of cell adhesion proteins at cellular adhesion sites, such as focal adhesions and adherens junctions.17-19 Consequently, polymeric cell substrates with immobilized linkers are not well suited to…

…biological processes, as well as physical, mechanical or chemical mechanisms is fluorescence microscopy that detects the universal luminescence family of processes in which susceptible molecules emit light from electronically excited states. Excitation of a…

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