title: Differential expression of living mammary epithelial cell subpopulations in milk during lactation in dairy cows.
abstract: Epithelial cells are shed into milk during lactation, and although they generally reflect the cellular characteristics of terminally differentiated luminal cells, previously the detection of more primitive cells was described in human milk where a cell population of epithelial lineage was detected expressing markers typical of progenitor cells. In this investigation, we report the development of flow cytometry analysis to allow multiparametric assessment of mammary epithelial cells observed in milk. Cells collected from milk samples of 10 healthy dairy cows were directly analyzed for 6 different markers: CD45, CD49f, cytokeratin 14, cytokeratin 18, presence of nucleus, and cell viability. Milk samples were collected in 3 different periods of lactation: early lactation (EL=d 0-30), mid-lactation (ML=d 90-120), and late lactation (LL=210-250). Here we identify the differential expression of precursor or differentiated cell markers (or both) in mammary epithelial cells present in bovine milk. Myoepithelial cells, as indicated by cells staining positively for cytokeratin 14(+)/cytokeratin 18(-), were observed to increase from EL to LL with a high correlation with nuclear staining inferring potential proliferative activity. Furthermore, a significant increase in CD49f(+) and cytokeratin 14(+)/cytokeratin 18(+) positive cells was observed in LL. This assay is a sensitive approach for evaluating the variations in the frequency and features of living epithelial cells, whose reciprocal balance may be significant in understanding mammary gland cellular function throughout lactation. These observations suggest that mammary epithelial cell immunophenotypes could be investigated as biomarkers for mammary gland function in dairy cows.
authors.affiliation: Department of Veterinary Sciences, Largo Braccini 2, University of Torino, 10095 Grugliasco (TO), Italy. Electronic address: mario.baratta@unito.it.
authors.forename: M
authors.initials: M
authors.lastname: Baratta
authors.affiliation: Department of Veterinary Sciences, Largo Braccini 2, University of Torino, 10095 Grugliasco (TO), Italy.
authors.forename: M G
authors.initials: MG
authors.lastname: Volpe
authors.affiliation: Department of Agricultural, Forest and Food Sciences, University of Torino, 10095 Grugliasco (TO), Italy.
authors.forename: D
authors.initials: D
authors.lastname: Nucera
authors.affiliation: Department of Comparative Biomedicine and Food Science, University of Padova, 35020 Legnaro (PD), Italy.
authors.forename: G
authors.initials: G
authors.lastname: Gabai
authors.affiliation: Department of Comparative Biomedicine and Food Science, University of Padova, 35020 Legnaro (PD), Italy.
authors.forename: N
authors.initials: N
authors.lastname: Guzzo
authors.forename: M
authors.initials: M
authors.lastname: Fustini
authors.affiliation: Department of Veterinary Medical Sciences, University of Bologna, 40064 Ozzano Emilia (BO), Italy.
authors.forename: M
authors.initials: M
authors.lastname: Faustini
authors.affiliation: Department of Veterinary Sciences, Largo Braccini 2, University of Torino, 10095 Grugliasco (TO), Italy.
authors.forename: E
authors.initials: E
authors.lastname: Martignani
mesh_terms: D000818:Animals; D002417:Cattle / Q000502:physiology*; D002452:Cell Count / Q000662:veterinary; D002454:Cell Differentiation; D002470:Cell Survival; D004847:Epithelial Cells / Q000145:classification / Q000276:immunology / Q000502:physiology; D005260:Female; D016130:Immunophenotyping / Q000662:veterinary; D007774:Lactation; D008321:Mammary Glands, Animal / Q000166:cytology* / Q000378:metabolism; D008892:Milk / Q000166:cytology* / Q000276:immunology* / Q000378:metabolism
keywords: bovine; cell precursor; epithelial cell; milk