MULTIPARAMETER ANALYSIS OF PRIMARY EPITHELIAL CULTURES GROWN ON CYCLOPORE MEMBRANES

The use of porous membranes as culture support for epithelial cells ha s previously been shown to cause functional differentiation of these c ells mimicking an in vivo condition, in contrast to culture on plastic . The different materials of which the membranes are made also have di fferent properties, such as transparency, rigidity, and retention of m olecules. Cyclopore membranes (polyethylene terephtalate) are permeabl e, transparent, rigid, and have low protein retention. In this study w e examined the applicability of assessing multiple parameters on a sin gle culture of primary epithelial cells on a Cyclopore membrane. Cultu res of transitional epithelial cells on these membranes differentiate into an organoid-like epithelium. We were able to perform morphometric analysis during and after cell culture and to quantitate proliferatio n and differentiation by double immuno-enzymatic staining. On these cu ltures, quantitative radiochemical analysis could also be achieved, re taining the morphology and the immunohistochemical staining. Cross-sec tions of paraffin-embedded and plastic-embedded cultures were analyzed qualitatively by light and transmission electron microscopy, respecti vely. Finally, cytokeratins in these cultures could also be visualized by immunofluorescence analysis. This suitability for simultaneous ass essment of both qualitative and quantitative parameters on a single ce ll culture grown on a Cyclopore membrane reduces the need of biologica l materials and may lead to better insight into physiological processe s.