Separation and characterization of basal and secretory cells from the rat trachea by flow cytometry

Abstract

Basal and secretory cells have been separated as highly enriched viable populations from single‐cell suspensions of rat tracheal epithelial cells. Isolation of the populations was achieved by preparation of a cell suspension and separation by flow cytometry using contour maps generated from 2° and 90° light scatter signals. Flow cytometric analysis of cells showed 10% of the whole preparation were cells in SG₂M phase of the cell cycle. The secretory cells accounted for 86% of these cycling cells; the remainder were accounted for by the basal cells. Culture of sorted populations of basal and secretory cells in serum free defined medium showed that basal cells had a lower (0.6%) colony‐forming efficiency than secretory cells (3.4%). Significant differences in blue auto‐fluorescence, Hoechst 33342 uptake, and lectin staining were apparent between basal and secretory cells. These results suggest that the secretory cell rather than the basal cell is primarily the cell type involved in maintenance of the normal tracheal epithelium. Secretory cells are greater in number, have a higher proliferative potential, and greater metabolic capability. Because of these traits they may be a critical cell at risk from damage by environmental agents.