A Fluid Membrane-Based Soluble Ligand Display System for Live CellAssays

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Cell communication modulates numerous biological processes including proliferation, apoptosis, motility, invasion and differentiation. Correspondingly, there has been significant interest in the development of surface display strategies for the presentation of signaling molecules to living cells. This effort has primarily focused on naturally surface-bound ligands, such as extracellular matrix components and cell membranes. Soluble ligands (e.g. growth factors and cytokines) play an important role in intercellular communications, and their display in a surface-bound format would be of great utility in the design of array-based live cell assays. Recently, several cell microarray systems that display cDNA, RNAi, or small molecules in a ... continued below

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Nam, Jwa-Min; Nair, Pradeep N.; Neve, Richard M.; Gray, Joe W. & Groves, Jay T. October 14, 2005.

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Cell communication modulates numerous biological processes including proliferation, apoptosis, motility, invasion and differentiation. Correspondingly, there has been significant interest in the development of surface display strategies for the presentation of signaling molecules to living cells. This effort has primarily focused on naturally surface-bound ligands, such as extracellular matrix components and cell membranes. Soluble ligands (e.g. growth factors and cytokines) play an important role in intercellular communications, and their display in a surface-bound format would be of great utility in the design of array-based live cell assays. Recently, several cell microarray systems that display cDNA, RNAi, or small molecules in a surface array format were proven to be useful in accelerating high-throughput functional genetic studies and screening therapeutic agents. These surface display methods provide a flexible platform for the systematic, combinatorial investigation of genes and small molecules affecting cellular processes and phenotypes of interest. In an analogous sense, it would be an important advance if one could display soluble signaling ligands in a surface assay format that allows for systematic, patterned presentation of soluble ligands to live cells. Such a technique would make it possible to examine cellular phenotypes of interest in a parallel format with soluble signaling ligands as one of the display parameters. Herein we report a ligand-modified fluid supported lipid bilayer (SLB) assay system that can be used to functionally display soluble ligands to cells in situ (Figure 1A). By displaying soluble ligands on a SLB surface, both solution behavior (the ability to become locally enriched by reaction-diffusion processes) and solid behavior (the ability to control the spatial location of the ligands in an open system) could be combined. The method reported herein benefits from the naturally fluid state of the supported membrane, which allows surface-linked ligands to diffuse freely in two dimensions. Ligands can become reorganized beneath cells, by reaction-diffusion processes, and may also adopt spatial configurations reflecting those of their cognate receptors on the cell surface (Figure 1B). This provides a significant benefit over conventional cell signaling and culturing systems that present inflexible distributions of signaling molecules. In this study, we observe marked differences in the response of cells to membrane surface displayed soluble ligands as a function of membrane fluidity. Tethering of soluble signaling molecules to fluid supported membranes opens up opportunities to use already developed membrane fabrication technologies to present soluble components within a surface array format.

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  • Journal Name: ChemBioChem; Journal Volume: 7; Journal Issue: 3; Related Information: Journal Publication Date: 03/2006

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  • Report No.: LBNL--59132
  • Grant Number: DE-AC02-05CH11231
  • Office of Scientific & Technical Information Report Number: 891352
  • Archival Resource Key: ark:/67531/metadc876554

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  • October 14, 2005

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  • Sept. 21, 2016, 2:29 a.m.

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  • Dec. 16, 2016, 1:07 p.m.

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Nam, Jwa-Min; Nair, Pradeep N.; Neve, Richard M.; Gray, Joe W. & Groves, Jay T. A Fluid Membrane-Based Soluble Ligand Display System for Live CellAssays, article, October 14, 2005; Berkeley, California. (digital.library.unt.edu/ark:/67531/metadc876554/: accessed July 18, 2018), University of North Texas Libraries, Digital Library, digital.library.unt.edu; crediting UNT Libraries Government Documents Department.