Epithelial cells in 3D cultures
Epithelial cells serve as a barrier between the body interior and the external environment. To accomplish this function, epithelial tissues maintain special characteristics such as a polarized morphology, unique cell-cell contacts and intimate interactions with the extracellular matrix (ECM). To model epithelia in vivo, epithelial cells such as MDCK can be grown embedded in a three dimensional ECM (3D cultures). MDCK cells grown in 3D cultures form spherical cysts characterized by a hollow lumen surrounded by polarized cells (see below). This ‘in vivo - like’ system recapitulates many features of epithelial tissues in vivo that are partly lost in the absence of ECM. Growth of MDCK cells in 3D cultures thus provides a unique opportunity to study the development and maintenance of epithelial cell polarity in a system that is readily amenable to experimental manipulation and detailed microscopic analyses under relatively physiological conditions. We are employing this system to study various aspects of epithelial cell function, polarity and cell-cell junctions. One current focus of study is the role of dynamin - dependent endocytosis in 3D culture formation, including the generation of cellular polarity and cell-cell junctions. This is being approached using both indirect immunofluorescence and live cell imaging.
Figure: Confocal cross section of MDCK cells grown in 3D culture. MDCK cells were grown embedded in collagen I for 5 days. The apical surface is labeled with Texas-red phalloidin (red), the basolateral membrane is stained with the adherent junction marker E-cadherin (green) and the nuclei are stained with Hoechst (blue).
For more information, contact Natalie Elia.
