title: Quantification of regenerative potential in primary human mammary epithelial cells.
abstract: We present an organoid regeneration assay in which freshly isolated human mammary epithelial cells are cultured in adherent or floating collagen gels, corresponding to a rigid or compliant matrix environment. In both conditions, luminal progenitors form spheres, whereas basal cells generate branched ductal structures. In compliant but not rigid collagen gels, branching ducts form alveoli at their tips, express basal and luminal markers at correct positions, and display contractility, which is required for alveologenesis. Thereby, branched structures generated in compliant collagen gels resemble terminal ductal-lobular units (TDLUs), the functional units of the mammary gland. Using the membrane metallo-endopeptidase CD10 as a surface marker enriches for TDLU formation and reveals the presence of stromal cells within the CD49f(hi)/EpCAM(-) population. In summary, we describe a defined in vitro assay system to quantify cells with regenerative potential and systematically investigate their interaction with the physical environment at distinct steps of morphogenesis.
authors.affiliation: Institute of Stem Cell Research, Helmholtz Center for Health and Environmental Research Munich, Neuherberg 85764, Germany.
authors.forename: Jelena R
authors.initials: JR
authors.lastname: Linnemann
authors.affiliation: Institute of Stem Cell Research, Helmholtz Center for Health and Environmental Research Munich, Neuherberg 85764, Germany.
authors.forename: Haruko
authors.initials: H
authors.lastname: Miura
authors.affiliation: Institute of Stem Cell Research, Helmholtz Center for Health and Environmental Research Munich, Neuherberg 85764, Germany.
authors.forename: Lisa K
authors.initials: LK
authors.lastname: Meixner
authors.affiliation: Institute of Experimental Genetics, Helmholtz Center Munich, Neuherberg 85764, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Irmler
authors.affiliation: Institute of Stem Cell Research, Helmholtz Center for Health and Environmental Research Munich, Neuherberg 85764, Germany.
authors.forename: Uwe J
authors.initials: UJ
authors.lastname: Kloos
authors.affiliation: Institute of Stem Cell Research, Helmholtz Center for Health and Environmental Research Munich, Neuherberg 85764, Germany.
authors.forename: Benjamin
authors.initials: B
authors.lastname: Hirschi
authors.affiliation: Institute of Pathology, Medical School, Ludwig Maximilian University Munich, Munich 80337, Germany.
authors.forename: Harald S
authors.initials: HS
authors.lastname: Bartsch
authors.affiliation: Institute of Computational Biology, Helmholtz Center Munich, Neuherberg 85764, Germany.
authors.forename: Steffen
authors.initials: S
authors.lastname: Sass
authors.affiliation: Institute of Experimental Genetics, Helmholtz Center Munich, Neuherberg 85764, Germany Department of Experimental Genetics, Technical University Munich, Freising 85354, Germany.
authors.forename: Johannes
authors.initials: J
authors.lastname: Beckers
authors.affiliation: Institute of Computational Biology, Helmholtz Center Munich, Neuherberg 85764, Germany Department of Mathematics, Technical University Munich, Garching 85747, Germany.
authors.forename: Fabian J
authors.initials: FJ
authors.lastname: Theis
authors.affiliation: Nymphenburg Clinic for Plastic and Aesthetic Surgery, Munich 80637, Germany.
authors.forename: Christian
authors.initials: C
authors.lastname: Gabka
authors.affiliation: Institute of Pathology, Medical School, Ludwig Maximilian University Munich, Munich 80337, Germany.
authors.forename: Karl
authors.initials: K
authors.lastname: Sotlar
authors.affiliation: Institute of Stem Cell Research, Helmholtz Center for Health and Environmental Research Munich, Neuherberg 85764, Germany christina.scheel@helmholtz-muenchen.de.
authors.forename: Christina H
authors.initials: CH
authors.lastname: Scheel
mesh_terms: D015415:Biomarkers / Q000378:metabolism*; D018929:Cell Culture Techniques / Q000379:methods*; D002469:Cell Separation / Q000379:methods; D003094:Collagen; D005260:Female; D005455:Fluorescent Antibody Technique; D020869:Gene Expression Profiling; D006801:Humans; D007201:Indicator Dilution Techniques; D042361:Mammary Glands, Human / Q000166:cytology* / Q000502:physiology*; D009024:Morphogenesis / Q000502:physiology*; D015260:Neprilysin / Q000378:metabolism; D009940:Organoids / Q000502:physiology*; D012038:Regeneration / Q000502:physiology*
chemical_list: D015415:Biomarkers; D003094:Collagen; D015260:Neprilysin
keywords: Basal; Branching morphogenesis; CD10; Collagen gel; Luminal; Mammary stem cell; Organoid; Primary human mammary epithelial cells; Progenitor cells; Regenerative potential; Terminal ductal-lobular units