title: Single cell 3-D platform to study ligand mobility in cell-cell contact.
abstract: Lateral mobility and dimensionality have both been shown to influence cellular behavior, but have yet to be combined and applied in a single in vitro platform to address, e.g., cell adhesion in a setting mimicking the three-dimensional environment of neighboring cells in a reductionist way. To study the effect of the lateral mobility of cell adhesive ligands in three dimensions we present and characterize a platform, which enables patterning of single cells into microwells presenting a cell membrane mimetic interface pre-patterned to its walls. Soluble E-cadherin extracellular domains coupled through an optimized streptavidin-antibody linkage to lipids in a supported lipid bilayer (SPB) were presented on the microwell walls as either laterally mobile or immobile ligands. The fluidity was controlled through a small change in temperature by choosing phospholipids for the SPB with a lipid phase transition temperature around 30 °C. The platform thus enabled the investigation of cell adhesion to either laterally immobile or mobile E-cadherin ligands presented on the same cell membrane mimetic surface. Chinese hamster ovary (CHO) cells engineered to express E-cadherin that were cultured on the platform demonstrated that enhanced cadherin lateral mobility significantly decreased the formation of actin bundles and resulted in more diffuse actin organization, while constraining the cell shape to that of the microwell. This example highlights the potential to use in vitro cell culture platforms to mimic direct cell-cell interaction in a controlled environment that nevertheless captures the dynamic nature of the native cell environment.
authors.affiliation: Laboratory for Surface Science and Technology, Department of Materials, ETH Zurich, 8093 Zurich, Switzerland.
authors.forename: Mirjam
authors.initials: M
authors.lastname: Andreasson-Ochsner
authors.forename: Gregory
authors.initials: G
authors.lastname: Romano
authors.forename: Maria
authors.initials: M
authors.lastname: Håkanson
authors.forename: Michael L
authors.initials: ML
authors.lastname: Smith
authors.forename: Deborah E
authors.initials: DE
authors.lastname: Leckband
authors.forename: Marcus
authors.initials: M
authors.lastname: Textor
authors.forename: Erik
authors.initials: E
authors.lastname: Reimhult
mesh_terms: D000818:Animals; D000906:Antibodies / Q000737:chemistry / Q000276:immunology; D001710:Biotin / Q000737:chemistry / Q000378:metabolism; D016466:CHO Cells; D015820:Cadherins / Q000737:chemistry / Q000378:metabolism; D002448:Cell Adhesion; D002450:Cell Communication; D016764:Cell Polarity; D006224:Cricetinae; D003412:Cricetulus; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D008024:Ligands; D008051:Lipid Bilayers / Q000737:chemistry; D019809:Streptavidin / Q000737:chemistry / Q000378:metabolism
chemical_list: D000906:Antibodies; D015820:Cadherins; D004129:Dimethylpolysiloxanes; D008024:Ligands; D008051:Lipid Bilayers; C013830:baysilon; D001710:Biotin; D019809:Streptavidin