LLC-PK1 cysts: a model for the study of epithelial polarity

In the present work, we have taken advantage of the properties of two recently isolated clonal subpopulations of the pig kidney-derived LLC-PK1 cell line to study aspects of the establishment of epithelial polarity. When grown in suspension, LLC-PK1/D + Sc cells reaggregated within a few hours and, during the following days of culture, formed free-floating, hollow spheres or cysts, lined by a monolayer of polarized cells. In contrast, LLC-PK1/D- cells were unable to develop such polarized structures even upon prolonged culture in suspension. The polarity of the LLC-PK1/D + Sc cells lining the cysts was inverted compared to that in intact renal tubules, the microvilli-rich "apical" pole being oriented toward the external medium. However, upon embedding these preformed cysts in collagen gels, a reversal of polarity was observed within hours, the microvilli-rich pole now facing the cyst cavity. Thus, in the same clonally derived cell population, cell-to-cell contact and interaction with the extracellular matrix differentially affect the orientation of cellular polarity. The LLC-PK1/D + Sc cysts provide a suitable in vitro model system for further study of the sequential events by which extracellular matrix components induce an appropriately oriented polarization. In addition, the comparison between LLC-PK1/D + Sc and D- cells, which differ in their ability to polarize in response to cell-to-cell contact, should help define some of the cellular determinants involved in epithelial organization.