Cell proliferation in bovine mammary epithelium: identification of the primary proliferative cell population

Histologic analyses indicate that a lightly staining cell population present in mammary parenchyma may function as mammary stem cells. We performed an analysis of mammary epithelial cell proliferation in prepubertal bromodeoxyuridine (BrdU)-injected Holstein heifers to investigate this hypothesis. We observed light, dark, and intermediate staining cells in histologic sections stained with basic fuschin and azure II. Light cells comprised 10% of the total parenchymal cell population but accounted for 50% of the cell proliferation. Intermediate cells comprised 60% of the cell population and 43% of proliferating cells. Dark cells comprised 30% of the parenchymal cell population but only 7% of proliferating cells. The distribution of BrdU+ cells across basal, embedded, and lumenal parenchymal cell layers was correlated with the fraction of total parenchymal cells present in each layer (r=0.99). However, the proportion of mitotic cells observed in the basal cell layer was only half of what would be predicted by the BrdU-labeling data. This observation suggests that some basal cells either arrest in G(2) or migrate into the suprabasal epithelial layers before undergoing mitosis. These results strongly support the concept that lightly staining mammary parenchymal cells are the primary proliferative cell population.