TY - JOUR
T1 - Mammospheres from murine mammary stem cell-enriched basal cells
T2 - Clonal characteristics and repopulating potential
AU - Dong, Qiaoxiang
AU - Wang, Danhan
AU - Bandyopadhyay, Abhik
AU - Gao, Hui
AU - Gorena, Karla M.
AU - Hildreth, Kim
AU - Rebel, Vivienne I.
AU - Walter, Christi A.
AU - Huang, Changjiang
AU - Sun, Lu Zhe
N1 - Funding Information:
This work was supported in part by funding from NIH Grants R01CA75253 and R01ES022057 , the Bank of America Shelby Rae Tengg Foundation , the Mary Kay Foundation (# 082-12 ), and the Cancer Therapy and Research Center at University of Texas Health Science Center at San Antonio through the NCI Cancer Center Support Grant 2P30CA054174-17 to the flow cytometry core facility. We thank Dr. John Stingl for his kind instruction in establishing the in vitro and in vivo assays in our laboratory. We also thank Dr. Rong Li for the use of the Nikon BioStation-IM.
PY - 2013
Y1 - 2013
N2 - 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.
AB - 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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U2 - 10.1016/j.scr.2013.01.007
DO - 10.1016/j.scr.2013.01.007
M3 - Article
C2 - 23466563
AN - SCOPUS:84874715117
SN - 1873-5061
VL - 10
SP - 396
EP - 404
JO - Stem Cell Research
JF - Stem Cell Research
IS - 3
ER -