RESUMO
These studies quantified the relative effects of E-cadherin expression and homophilic ligation on the integrin-mediated motility of epithelial cells. Micropatterned proteins were used to quantitatively titrate the ligation of E-cadherin and integrin receptors in order to assess their coordinate influence on the migration velocities of MDA-MB-231 breast tumor epithelial cells. Fibronectin, E-cadherin, and mixtures of fibronectin and E-cadherin were covalently patterned on solid surfaces at defined compositions and mass coverages. The migration velocities of parental epithelial cells and of cells engineered to express E-cadherin under tetracycline control show that E-cadherin expression reduces cell motility by both adhesion-dependent and adhesion-independent mechanisms. Increasing E-cadherin expression levels also suppresses the dependence of cell velocity on the fibronectin coverage. On E-cadherin-containing substrata, the cell velocity decreases both with the E-cadherin expression level and with the immobilized E-cadherin surface density. These studies thus identified conditions under which E-cadherin preferentially suppresses cell migration by adhesion-independent versus adhesion-dependent mechanisms.
Assuntos
Caderinas/química , Células Epiteliais/metabolismo , Integrinas/química , Caderinas/metabolismo , Adesão Celular , Linhagem Celular Tumoral , Movimento Celular , Células Epiteliais/citologia , Fibronectinas/química , Fibronectinas/metabolismo , Citometria de Fluxo/métodos , Humanos , Proteínas/química , Ressonância de Plasmônio de Superfície , Propriedades de Superfície , Engenharia Tecidual/métodosRESUMO
This paper explores the effects of the surface density and concentration profiles of extra cellular matrix proteins on the migration of rat intestinal IEC-6 cells. Microfluidic devices were used to create linear, immobilized gradients of laminin. This study investigated both the impact of the steepness and local concentrations on the directedness of cell migration. The bulk concentrations of proteins in the feed streams in the mixing device determined the gradient profile and the local concentration of laminin in the device. Two sets of gradients were used to explore cell migration directedness: (i) gradients with similar change in local concentration, i.e., the same gradient steepness, and (ii) different gradients with similar local concentrations. Cells migrated up the gradients, independent of the steepness of the gradients used in this study. At the same local laminin concentration, the migration rate was independent of the gradient steepness. However, cell directedness decreased significantly at high laminin densities.
Assuntos
Movimento Celular/fisiologia , Polaridade Celular/fisiologia , Proteínas da Matriz Extracelular/química , Actinas/metabolismo , Animais , Linhagem Celular , Técnicas In Vitro , Intestino Delgado/citologia , Intestino Delgado/metabolismo , Laminina/química , Microfluídica , Ratos , Ressonância de Plasmônio de Superfície , Propriedades de Superfície , Vinculina/metabolismoRESUMO
This work describes the use of microfluidic tools to generate covalently immobilized counter gradients of extracellular matrix (ECM) proteins laminin and collagen I. Using these platforms, we demonstrate control of the expression levels of two proteins linked to cell cycle progression by virtue of the spatial location of cells on the gradients, and hence by the local ECM environments in these devices. In contrast to physisorbed gradients, covalently immobilized protein patterns preserved the gradient fidelity, making long term cell studies feasible. This method of precisely controlling local cell environments is simple and broadly portable to other cell types and to other ECM proteins or soluble factors. Our approach promises to enable new investigations in cell biology that will contribute to the establishment of biological design rules for controlling cell growth, differentiation, and function.