期刊
DEVELOPMENT
卷 144, 期 6, 页码 1065-1071出版社
COMPANY OF BIOLOGISTS LTD
DOI: 10.1242/dev.145045
关键词
3D imaging; Mammary organoids; Clonality; In vitro lineage tracing; Multicolor reporter mice; Stem/progenitor cells
资金
- Australian National Health and Medical Research Council (NHMRC) [1016701, 1085191, 1086727]
- NHMRC Independent Research Institute Infrastructure Support Scheme (IRIISS)
- Victorian State Government through Victorian Cancer Agency
- Australian Cancer Research Foundation
- Marie Sklodowska Curie global individual fellowship of the European Commission
- National Breast Cancer Foundation (NBCF)/Cure Cancer Australia Foundation
- NHMRC [1078730, 1037230, 1102742]
- National Health and Medical Research Council of Australia [1078730, 1102742, 1086727, 1085191] Funding Source: NHMRC
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 formation in vitro. Here, 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 lineagespecific reporter mice established the bipotent capacity of basal cells and the restricted potential of luminal cells. In summary, we describe optimized in vitro conditions 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.
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