title: Clonogenic assay allows for selection of a primitive mammary epithelial cell population in bovine.
abstract: Adult mammary stem cells have been identified in several species including the bovine. They are responsible for the development of the gland and for cyclic remodeling during estrous cycles and pregnancy. Epithelial cell subpopulations exist within the mammary gland. We and others showed previously that the Colony Forming Cell (CFC) assay can be used to detect lineage-restricted mammary progenitors. We carried out CFCs with bovine mammary cells and manually separated colonies with specific morphologies associated with either a luminal or a myoepithelial phenotype. Expression of specific markers was assessed by immunocytochemistry or by flow cytometry to confirm that the manual separation resulted in isolation of phenotipically different cells. When transplanted in recipient immunodeficient mice, we found that only myoepithelial-like colonies gave rise to outgrowths that resembled bovine mammary alveoli, thus proving that adult stem cells were maintained during culture and segregated with myoepithelial cells. After recovery of the cells from the transplanted mice and subsequent progenitor content analysis, we found a tendency to detect a higher progenitor frequency when myoepithelial-like colonies were transplanted. We here demonstrate that bovine adult mammary stem cells can be sustained in short-term culture and that they can be enriched by manually selecting for basal-like morphology.
authors: Department of Veterinary Science, University of Turin, Largo Braccini 2, 10095 Grugliasco, TO, Italy. Electronic address: eugenio.martignani@unito.it.
authors: Eugenio
authors: E
authors: Martignani
authors: Department of Veterinary Science, University of Turin, Largo Braccini 2, 10095 Grugliasco, TO, Italy. Electronic address: diego.cravero@unito.it.
authors: Diego
authors: D
authors: Cravero
authors: Department of Veterinary Science, University of Turin, Largo Braccini 2, 10095 Grugliasco, TO, Italy. Electronic address: silvia.miretti@unito.it.
authors: Silvia
authors: S
authors: Miretti
authors: Department of Veterinary Science, University of Turin, Largo Braccini 2, 10095 Grugliasco, TO, Italy. Electronic address: paolo.accornero@unito.it.
authors: Paolo
authors: P
authors: Accornero
authors: Department of Veterinary Science, University of Turin, Largo Braccini 2, 10095 Grugliasco, TO, Italy. Electronic address: mario.baratta@unito.it.
authors: Mario
authors: M
authors: Baratta
mesh_terms: D000818:Animals; D001681:Biological Assay / Q000379:methods; D015415:Biomarkers / Q000378:metabolism; D002417:Cattle; D004847:Epithelial Cells / Q000166:cytology*; D005260:Female; D006801:Humans; D008321:Mammary Glands, Animal / Q000166:cytology*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D010641:Phenotype; D013234:Stem Cells / Q000166:cytology*
chemical_list: D015415:Biomarkers
keywords: Bovine; Epithelial Cell; Mammary gland; Stem cell; Xenotransplantation