title: Mobile and three-dimensional presentation of adhesion proteins within microwells.
abstract: On traditional cell culture substrates cells adhere to a planar 2D surface where ligands are presented immobile. A more realistic presentation of cell adhesion ligands which can account for lateral mobility and a more tissue-like 3D presentation would allow studies addressing fundamental questions of significant importance for applications such as tissue engineering and implant intregration. To study the effect of lateral mobility of cell membrane interaction cues in three dimensions, we have developed and characterized a platform which generically enables patterning of single cells into microwells presenting a cell membrane mimetic interface pre-patterned to its walls. Here, we describe its application in presenting a soluble cell adhesive ligand coupled through streptavidin-antibody linkage to lipids in a supported lipid bilayer (SLB) coated microwell. The lateral mobility of the presented ligands was controlled through a small change in temperature. The SLB phospholipid composition was choosen such that below its melting transition at 30 °C the ligands are immobile, while above 30 °C they are laterally mobile. The platform thus enables the investigation of cell adhesion to either laterally immobile or mobile E-cadherin ligand presented on the same cell membrane mimetic surface.
authors.affiliation: Department of Materials, Laboratory for Surface Science and Technology, ETH Zurich, Zurich, Switzerland.
authors.forename: Mirjam
authors.initials: M
authors.lastname: Andreasson-Ochsner
authors.forename: Erik
authors.initials: E
authors.lastname: Reimhult
mesh_terms: D000906:Antibodies / Q000737:chemistry; D015820:Cadherins / Q000378:metabolism; D002448:Cell Adhesion / Q000502:physiology*; D018929:Cell Culture Techniques / Q000379:methods*; D002462:Cell Membrane / Q000737:chemistry* / Q000276:immunology; D006801:Humans; D008024:Ligands; D008051:Lipid Bilayers / Q000737:chemistry / Q000378:metabolism; D008967:Molecular Biology / Q000379:methods*; D019809:Streptavidin / Q000737:chemistry
chemical_list: D000906:Antibodies; D015820:Cadherins; D008024:Ligands; D008051:Lipid Bilayers; D019809:Streptavidin