title: Development of a Model System to Study Cell Adhesion and Cell Mechanics
creator: Eberhard, Christian
subject: ddc-500
subject: 500 Natural sciences and mathematics
subject: ddc-530
subject: 530 Physics
subject: ddc-570
subject: 570 Life sciences
description: Within this work, a simplified model system was developed for the experimental investigation of physical mechanisms underlying cell adhesion. Cellular complexity was reduced to three essential components: the lipid bilayer, the cell adhesion molecule integrin and the cytoskeletal protein actin. Integrin was integrated in spherical lipid bilayers, called giant unilamellar vesicles (GUVs), which were additionally filled with actin.  Integrin alphaIIb beta3 was purified from human platelet membranes with high yield. Initially, this integrin was reconstituted in small unilamellar vesicles (SUVs). By variation of lipid composition, vesicle size and detergent concentration, crucial parameters for efficient integration could be determined and incorporation efficiency was enhanced.  Two different approaches were compared to grow GUVs: electroformation and spontaneous swelling on agarose-lipid films. Besides biochemical characterization, formation of GUVs was investigated by means of confocal laser scanning microscopy confirming successful integration of integrin into the bilayer of the giant vesicles.  A novel method for the preparation of GUVs from purified platelet membranes was established. Platelet membranes were directly purified and grown to giant vesicles by electroformation. Interaction of these GUVs with nano-structured and bio-functionalized surfaces was investigated. Adhesion of these integrin-coated vesicles was characterized for the first time by reflection interference contrast microscopy (RICM), which allowed estimation of an average adhesion energy.
date: 2012-12-11
type: Dissertation
type: info:eu-repo/semantics/doctoralThesis
type: NonPeerReviewed
format: application/pdf
identifier: https://archiv.ub.uni-heidelberg.de/volltextserverhttps://archiv.ub.uni-heidelberg.de/volltextserver/14257/1/PhD_Thesis_Christian_Eberhard.pdf
identifier: DOI:10.11588/heidok.00014257
identifier: urn:nbn:de:bsz:16-heidok-142576
identifier:   Eberhard, Christian  (2012) Development of a Model System to Study Cell Adhesion and Cell Mechanics.  [Dissertation]     
relation: https://archiv.ub.uni-heidelberg.de/volltextserver/14257/
rights: info:eu-repo/semantics/openAccess
rights: http://archiv.ub.uni-heidelberg.de/volltextserver/help/license_urhg.html
language: eng