Mammospheres from murine mammary stem cell-enriched basal cells: Clonal characteristics and repopulating potential |
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Authors: | Qiaoxiang Dong Danhan Wang Abhik Bandyopadhyay Hui Gao Karla M Gorena Kim Hildreth Vivienne I Rebel Christi A Walter Changjiang Huang Lu-Zhe Sun |
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Institution: | 1. Department of Cellular & Structural Biology, University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78299, USA;2. School of Environmental Sciences and Public Health, Wenzhou Medical College, University Town, Wenzhou 325035, PR China;3. Cancer Therapy and Research Center, University of Texas Health Science Center, 7979 Wurzbach Road, San Antonio, TX 78299, USA;4. Flow Cytometry Core Facility, University of Texas Health Science Center, 7703 Floyd Curl Drive, San Antonio, TX 78299, USA;5. Greehey Children''s Cancer Research Institute, University of Texas Health Science Center, 8403 Floyd Curl Drive, San Antonio, TX 78299, USA;6. South Texas Veteran''s Health Care System, 7400 Merton Minter, San Antonio, TX 78229, USA |
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Abstract: | Identification of murine mammary stem cells (MaSCs) has been attempted with various in vitro and in vivo assays. While, the in vivo repopulation assay remains as the most definitive assay for MaSC detection, it is expensive, time-consuming, and technically challenging. The in vitro mammosphere assay was considered unreliable because of major concerns about its clonal origin. In the current study, co-culture experiments with mammary cells from fluorescent protein transgenic mice and time-lapse video microscopy revealed that > 90% mammospheres formed from sorted basal epithelial-enriched cells were of clonal origin in terms of stem cell. These basal-cell derived mammospheres were further distinguished morphologically in a 3-dimensional extracellular matrix culture and functionally in the in vivo repopulation assay. Transplant of single mammospheres or the resultant 3-dimensional solid structures into gland-free mammary fat pads yielded a 70% success rate of multilineage mammary gland reconstitution. Thus, this in vitro sphere formation and differentiation assay is a reliable alternative to the in vivo repopulation assay for the study of MaSCs. |
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