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ORIGINAL RESEARCH ARTICLE

Identification of Putative Bovine Mammary Epithelial Stem Cells by Their Retention of Labeled DNA Strands

Bovine Functional Genomics Laboratory, USDA-ARS, Beltsville, Maryland 20705

To whom requests for reprints should be addressed at ¹ Bovine Functional Genomics Laboratory, USDA-ARS, Building 200, Room 14 Powder-Mill Road, BARC-East, Beltsville, MD 20705. E-mail: tony.capuco{at}ars.usda.gov

Stem cells appear to retain labeled DNA for extended periods because of their selective segregation of template DNA strands during mitosis. In this study, proliferating cells in the prepubertal bovine mammary gland were labeled using five daily injections of 5-bromo-2-deoxyuridine (BrdU). Five weeks later, BrdU-labeled mammary epithelial cells were still evident. The percentage of BrdU-labeled epithelial cells was greatest in the lower region of the mammary gland, near the gland cistern, and was decreased toward the periphery of the parenchymal region, where the ducts were invading the mammary fat pad. Increased numbers of BrdU-labeled epithelial cells in basal regions of the gland are likely a consequence of decreased proliferation rates and increased cell cycle arrest in this area. In peripheral regions of mammary parenchyma, the percentage of heavily labeled epithelial cells averaged 0.24%, a number that is consistent with estimates of the frequency of stem cells in the mouse mammary gland. Epithelial label-retaining cells seemingly represent a slowly proliferating population of cells, as 5.4% of heavily labeled cells were positive for the nuclear proliferation antigen Ki67. Because epithelial label-retaining cells contain estrogen receptor (ER)-negative and ER-positive cells, they apparently comprise a mixed population, which I suggest is composed of ER-negative stem cells and ER-positive progenitors. Continuing studies will address the usefulness of this technique to identify bovine mammary stem cells and to facilitate studies of stem cell biology.

Key Words: progenitor cells • estrogen receptor • mammary cell proliferation • label retaining cells • bromodeoxyuridine

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