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Mammary Epithelial Cell Lineage Changes During Cow’s Life

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Abstract

Milk production is highly dependent on the optimal development of the mammary epithelium. It is therefore essential to better understand mammary epithelial cell growth and maintenance from the related epithelial lineage during the animal life. Here, we characterized the epithelial lineage at puberty, lactation and dry-off in bovine using the cell surface markers CD49f, CD24, and CD10. The pubertal period was characterized by a high proportion of CD49fpos cells corresponding to various epithelial subpopulations, notably the CD24pos subpopulations. The proportion of CD49fpos cells was weaker during lactation and dry-off, and CD24pos cells were relatively few. Of note, the (sub)population profile at dry-off appeared close to that during lactation. Using a targeted gene approach, we associated specific genes with epithelial subpopulations, their expression level varying, or not, according to physiological stages. Caseins were only expressed in the CD49fmedCD24neg subpopulation. Basal marker genes (keratin(KRT)5, KRT14 and αSMA) were found in the CD49fhighCD24neg subpopulations. Luminal gene markers (KRT7, KRT8 and KRT19, CDH1 and the PRLR) were expressed in the CD49flowCD24neg subpopulation. The CD49flowCD24pos subpopulation, only abundant at puberty, expressed luminal gene markers and KI67 at high level. In contrast to others, the CD49fhighCD24pos cells accounted for a small proportion of total cells, decreasing from puberty to dry-off. They were characterized by expression of luminal and basal gene markers and low KI67 level. Interestingly, this subpopulation showed a remarkable stability of gene expression profile throughout physiological stages and bear the hallmark of quiescence that designate them as the potential bovine mammary stem cells.

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Acknowledgements

This work was supported by the Animal Physiology & Livestock System Department of the French National Institute for Agricultural Research (INRA). We thank Laurent Deleurme and Gersende Lacombe from the CytomeTRI platform as well as Marine Biget from the GEH platform of the BIOSIT core facility at Rennes (France) for technical assistance. Acknowledgements are also extended to the staff at the INRA dairy farm of Méjusseaume (UMR1348 PEGASE, Le Rheu, France); Sandra Wiart and Frédérique Mayeur-Nickel for laboratory analyses and Rémi Resmond for technical assistance in statistics.

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  1. PEGASE, INRA, Agrocampus Ouest , 35590, Saint-Gilles, France

    Laurence Finot, Eric Chanat & Frederic Dessauge

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  1. Laurence Finot
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  2. Eric Chanat
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  3. Frederic Dessauge
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Contributions

L. Finot and F. Dessauge designed and conceived experiments, L. Finot performed experiments, assemblied and analysed data. L. Finot, E.Chanat and F. Dessauge collaborated to the data interpretation. The three authors collaborated equally on the writing of the manuscript and its final approval.

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Correspondence to Frederic Dessauge.

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Figure S1

Populations of cells in the bovine mammary tissue highlighted by the expression of the surface proteins CD49f, CD24 or CD10 in flow cytometry. Dissociated cells from the mammary tissue of lactating and dried cows were stained with either anti-CD49f–FITC (CD49f), anti-CD24-APC (CD24) or anti-CD10-PE Vio770 (CD10) antibodies and analyzed by flow cytometry. Each gating shows the positive cells. Abbreviation: SSC, Side Scatter light (PDF 239 kb)

Figure S2

Sub-populations of epithelial cells in the bovine mammary epithelium highlighted by the co-expression of surface proteins CD49f, CD24 and CD10 in flow cytometry. Dissociated cells from the mammary tissue of lactating and dried cows were co-stained with either anti-CD49f-FITC and anti-CD10-PE Vio770 antibodies, or anti-CD10-PE Vio770 and anti-CD24-APC antibodies, and analyzed by flow cytometry. Each gating shows the positive cells; positive cells are located to the right of the gating on the x-axis and above the gating on the y-axis. Sub-populations of epithelial cells were distinguished according to the intensity of the cell surface marker expression (low vs. high). Abbreviation: SSC, Side Scatter light (PDF 218 kb)

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Finot, L., Chanat, E. & Dessauge, F. Mammary Epithelial Cell Lineage Changes During Cow’s Life. J Mammary Gland Biol Neoplasia 24, 185–197 (2019). https://doi.org/10.1007/s10911-019-09427-1

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