Methods for analysis of apical lumen trafficking using micropatterned 3D systems.

摘要: Epithelial organs are made of interconnected branched networks tubules, with a central lumen lined by monolayer epithelial cells. Certain cell lines can be converted into organotypic cultures the addition extracellular matrix components. When cultured in these conditions, cells reorient axis polarity, reorganize membrane surfaces, and transport apical proteins to form process that recapitulates essential aspects de novo formation during organ morphogenesis. Micropatterns simple technique allows culture controlled adhesive environment extremely high precision, close nanometer scale. We have recently developed method MDCK cysts on micropatterns different sizes composition. Using this we found changes micropattern shape size used modify contractility understand its contribution formation. imaging main advantage is apical-directed vesicle trafficking visualized x-y plane, which presents higher resolution confocal microscopes. Thus, use an efficient setup analyze polarized secretion unprecedented both time space.