Culture and characterization of epithelial cells from bovine choroid plexus

Abstract

Epithelial cells were isolated from choroid plexus, which plays a major role in cerebrospinal fluid production and regulation. Incubation of bovine choroid plexuses with pronase released cells which attached to plastic dishes with a plating efficiency of 5%. The cells were predominantly polygonal as judged by phase-contrast microscopy. These polygonal cells undergo limited cell division and survive for 1-2 weeks in culture before being overgrown by fibroblasts. The fibroblastic cells could be selectively removed from the cultures but the addition of 100 microgram/ml cis-hydroxyproline to the medium for several days. The specific activities of three membrane-bound enzymes, gamma-glutamyl transpeptidase, alkaline phosphatase, and leucine aminopeptidase were compared in selective cultures of polygonal cells and fibroblasts. Polygonal cells were found to have 4-5 times the gamma-glutamyl transpeptidase of fibroblasts, whereas fibroblasts have 2-3 times the alkaline phosphatase of polygonal cells. Leucine aminopeptidase levels in the two cultures were roughly equivalent. The polygonal cells rapidly lost gamma-glutamyl transpeptidase activity over a 4-day period in culture but acquired increased levels of leucine aminopeptidase. Alkaline phosphatase remained roughly constant. Under similar conditions fibroblasts showed a 3- to 4-fold increase in the specific activities of all three enzymes; these changes coincided with a substantial increase in cell density. Based on morphology, resistance to cis-hydroxyproline, absence of antihemophilic factor antigen, and enzymatic characteristics, we believe the polygonal cells to be of epithelial origin.