Scanning Electron Microscopy


We have used scanning electron microscopy (SEM) to examine the surface morphology of the renal epithelial cell lines MDCK and LLC-PK1 to determine the influence of alternative culture substrate conditions on cell polarity. We observed that regardless of physical culture conditions, cells establishes and maintained polarity, expressed by the characteristics of apical and basal surfaces. culture conditions did, however, influence the orientation of cell polarity in vitro. MDCK cells were grown within collagen gel, in which individual cells exhibited clonal growth to form fluid-filled epithelial cysts. The cells of MDCK-cysts were polarized with apical surface facing the lumen and basal surface against the surrounding collagen gel. This configuration made it possible to gain direct visual access, by SEM, to the basal surface by removing the supportive collagen lattice. The apical surface of MDCK-cysts was lined by short microvilli. Each cell possessed a solitary cilium. In comparison, the basal surface had few appendages, although cell boundaries were marked by interdigitating short processes. LLC-PK1 cells in monolayer culture bore solitary cilia and long microvilli at their apical surface. The basal surface of cells involved in dome formation was observed to possess only a sparse population of short, blunt processes. When LLC-PK1 cells were raised in stationary suspension culture or in monolayer atop non-culture grade plastic, they formed cysts with the cell apex facing the surrounding medium. These cells showed variable apical morphology. The cells of large, highly expanded cysts were often attenuated and had a relatively smooth apical surface. The basal surface of cells of fractured LLC-PK1 cysts commonly was also smooth, without prominent appendages.

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