title: Derivation of a robust mouse mammary organoid system for studying tissue dynamics.
abstract: Advances in stem cell research have enabled the generation of 'mini organs' or organoids that recapitulate phenotypic traits of the original biological specimen. Although organoids have been demonstrated for multiple organ systems, there are more limited options for studying mouse mammary gland formationin vitroHere, we have built upon previously described culture assays to define culture conditions that enable the efficient generation of clonal organoid structures from single sorted basal mammary epithelial cells (MECs). Analysis of Confetti-reporter mice revealed the formation of uni-colored structures and thus the clonal nature of these organoids. High-resolution 3D imaging demonstrated that basal cell-derived complex organoids comprised an inner compartment of polarized luminal cells with milk-producing capacity and an outer network of elongated myoepithelial cells. Conversely, structures generated from luminal MECs rarely contained basal/myoepithelial cells. Moreover, flow cytometry and 3D microscopy of organoids generated from lineage-specific reporter mice established the bipotent capacity of basal cells and the restricted potential of luminal cells. In summary, we describe optimizedin vitroconditions for the efficient generation of mouse mammary organoids that recapitulate features of mammary tissue architecture and function, and can be applied to understand tissue dynamics and cell-fate decisions.
authors.affiliation: Stem Cells and Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Paul R
authors.initials: PR
authors.lastname: Jamieson
authors.affiliation: Stem Cells and Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Johanna F
authors.initials: JF
authors.lastname: Dekkers
authors.affiliation: Stem Cells and Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Anne C
authors.initials: AC
authors.lastname: Rios
authors.affiliation: Stem Cells and Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Nai Yang
authors.initials: NY
authors.lastname: Fu
authors.affiliation: Stem Cells and Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia.|Familial Cancer Centre and Department of Medical Oncology, The Royal Melbourne Hospital, Parkville, Victoria 3050, Australia.|Department of Medicine, The University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Geoffrey J
authors.initials: GJ
authors.lastname: Lindeman
authors.affiliation: Stem Cells and Cancer Division, The Walter and Eliza Hall Institute of Medical Research, Parkville, Victoria 3052, Australia visvader@wehi.edu.au.|Department of Medical Biology, The University of Melbourne, Parkville, Victoria 3010, Australia.
authors.forename: Jane E
authors.identifier: 0000-0001-9173-6977
authors.initials: JE
authors.lastname: Visvader
mesh_terms: D000818:Animals; D019070:Cell Lineage; D002999:Clone Cells; D004847:Epithelial Cells / Q000166:cytology; D005260:Female; D005434:Flow Cytometry; D017930:Genes, Reporter; D021621:Imaging, Three-Dimensional; D008321:Mammary Glands, Animal / Q000166:cytology / Q000254:growth & development*; D051379:Mice; D018613:Microscopy, Confocal; D009940:Organoids / Q000166:cytology*; D046509:Tissue Culture Techniques / Q000379:methods*
keywords: 3D imaging; Clonality; In vitro lineage tracing; Mammary organoids; Multicolor reporter mice; Stem/progenitor cells