Abstract
Adult bovine mammary stem cells possess the ability to regenerate in vivo clonal outgrowths that mimic functional alveoli. Commonly available techniques that involve immunophenotype-based cell sorting yield cell fractions that are moderately enriched, far from being highly purified. Primary bovine mammary epithelial cells segregated in four different populations according to the expression of P-Cadherin and CD49f. Sorted cells from each fraction were tested for the presence of lineage-restricted progenitors and stem cells. Only cells from the CD49fhigh/P-Cadherinneg subpopulation were able to give rise to both luminal- and myoepithelial-restricted colonies in vitro and generate organized outgrowths in vivo, which are hallmarks of stem cell activity. After whole transcriptome analysis, we found gene clusters to be differentially enriched that relate to cell-to-cell communication, metabolic processes, proliferation, migration and morphogenesis. When we analyzed only the genes that were differentially expressed in the stem cell enriched fraction, clusters of downregulated genes were related to proliferation, while among the upregulated expression, cluster of genes related to cell adhesion, migration and cytoskeleton organization were observed. Our results show that P-Cadherin separates mammary subpopulations differentially in progenitor cells or mammary stem cells. Further we provide a comprehensive observation of the gene expression differences among these cell populations which reinforces the assumption that bovine mammary stem cells are typically quiescent.
Highlights
Adult bovine mammary stem cells possess the ability to regenerate in vivo clonal outgrowths that mimic functional alveoli
This study reports for the first time the utility that P-Cadherin may have in the purification of adult stem cells of the mammary gland in the bovine species
At the level of the mammary gland it has been reported that P-Cadherin is co-expressed with markers of the basal layer of the mammary alveolus in the mouse in association with CK5 and CD49f40 but it has not been investigated in more detail if it has a relevance for the regulation of the stem and / or progenitor cells
Summary
Adult bovine mammary stem cells possess the ability to regenerate in vivo clonal outgrowths that mimic functional alveoli. It has been well established that the cyclical development of the mammary gland in mammals represents a valuable model of the processes underlying cell differentiation for the dynamic remodeling of the organ and of its function, in this case, the production and secretion of milk This interest was further strengthened when in 2006 two studies showed the presence of a single pluripotent cell possessed the ability to completely regenerate the functional unit of the gland, the mammary alveolus[1,2]. P-Cadherin is a calcium-dependent cell–cell adhesion glycoprotein with a homeostatic function in several tissues and its name is derived from the site where this adhesion molecule was firstly characterized, the placenta[18] It exerts a crucial role in the conservation of the structural integrity of epithelial tissues and regulates processes involved in embryonic development and maintenance of adult tissue architecture, important for cell differentiation, cell shape, cell polarity, growth and migration[19,20,21]
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