Development of a Novel Single-Cell Attachment and Spreading Platform Utilizing Fused-Fiber Nanonets

dc.contributor.authorGill, Amritpal Singhen
dc.contributor.committeechairNain, Amrinderen
dc.contributor.committeememberKapania, Rakesh K.en
dc.contributor.committeememberBehkam, Baharehen
dc.contributor.departmentMechanical Engineeringen
dc.date.accessioned2016-11-26T07:00:14Zen
dc.date.available2016-11-26T07:00:14Zen
dc.date.issued2015-06-04en
dc.description.abstractInitial attachment to the extracellular matrix (ECM) and consequent spreading is a necessary process in the cell cycle of which little is known. Cell spreading has been well-recognized in 2D systems, however, the native fibrous ECM presents cells with 3D biophysical cues. Thus, using suspended fibers as model systems, we present the development of a novel platform (Cell-STEPs) capable of capturing cell attachment dynamics and forces from the moment a cell in suspension contacts the fiber. Cell-STEPs comprises of a custom glass-bottom petri dish with a lid to deliver a constant supply of CO2 to maintain pH. Fibrous scaffolds are attached in the dish to allow cellular investigations over extended periods of time. We find that cell-fiber attachment occurs in three progressive phases: initial attachment of cell to fiber (phase 0), rapid drop in circularity (phase 1), and increase in cell spread area (phase 2). Furthermore, using iterative inverse methods, forces involved in cell spreading through deflection of fibers were estimated. Our findings provide new insights in attachment biomechanics, including initial sensing and latching of cell to fiber with a negligible or protrusive force, followed by rapid loss in circularity through protrusion sensing at nearly constant spread area and minimal force generation, transitioning to a final phase of increased contractile forces until spread area and force saturation is observed. Also, anisotropic spreading of cells on single and two-fibers are closely related, while cells attached to several fibers take longer and spread isotropically. The Cell-STEPs platform allows, for the first time, detailed interrogations in the discrete and orchestrated adhesion steps involved in cell-fibrous matrix recognition and attachment along with simultaneous measurements of forces involved in cell attachment.en
dc.description.degreeMaster of Scienceen
dc.format.mediumETDen
dc.identifier.othervt_gsexam:5651en
dc.identifier.urihttp://hdl.handle.net/10919/73504en
dc.publisherVirginia Techen
dc.rightsIn Copyrighten
dc.rights.urihttp://rightsstatements.org/vocab/InC/1.0/en
dc.subjectnanofiberen
dc.subjectattachmenten
dc.subjectcell forceen
dc.subjectcell spreadingen
dc.subjectsingle-cellen
dc.titleDevelopment of a Novel Single-Cell Attachment and Spreading Platform Utilizing Fused-Fiber Nanonetsen
dc.typeThesisen
thesis.degree.disciplineMechanical Engineeringen
thesis.degree.grantorVirginia Polytechnic Institute and State Universityen
thesis.degree.levelmastersen
thesis.degree.nameMaster of Scienceen
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