TY - JOUR
T1 - Stem/progenitor cells in murine mammary gland
T2 - Isolation and functional characterization
AU - Bandyopadhyay, Abhik
AU - Dong, Qiaoxiang
AU - Sun, Lu Zhe
PY - 2012
Y1 - 2012
N2 - The presence of adult functional mammary epithelial stem/progenitor cells in mammary gland and recent identification of enriched fraction of transplantable mammary stem cells (MaSCs) through specific cell surface markers have revolutionized the study to delineate the role of mammary stem/progenitor functions in both mammary gland development and mammary tumorigenesis. In this chapter, we have described detailed methods of isolation of mammary epithelial cells from murine mammary glands, enrichment of stem/progenitor fractions by fluorescence-activated cell sorting (FACS), in vitro mammosphere culture, and differentiation assays in two- and three-dimensional culture. We have also described the detailed protocol of in vivo cleared mammary fat pad transplantation assay for the assessment of the MaSC repopulating activity, which indicates their multilineage differentiation and self-renewal potential in vivo and is considered the "gold standard" assay of functional stem cells.
AB - The presence of adult functional mammary epithelial stem/progenitor cells in mammary gland and recent identification of enriched fraction of transplantable mammary stem cells (MaSCs) through specific cell surface markers have revolutionized the study to delineate the role of mammary stem/progenitor functions in both mammary gland development and mammary tumorigenesis. In this chapter, we have described detailed methods of isolation of mammary epithelial cells from murine mammary glands, enrichment of stem/progenitor fractions by fluorescence-activated cell sorting (FACS), in vitro mammosphere culture, and differentiation assays in two- and three-dimensional culture. We have also described the detailed protocol of in vivo cleared mammary fat pad transplantation assay for the assessment of the MaSC repopulating activity, which indicates their multilineage differentiation and self-renewal potential in vivo and is considered the "gold standard" assay of functional stem cells.
KW - Cleared mammary fat pad
KW - Fluorescence-activated cell sorting
KW - Mammary glands
KW - Mammary progenitor cells
KW - Mammary stem cells
KW - Mammosphere
KW - Mice
UR - http://www.scopus.com/inward/record.url?scp=84861808502&partnerID=8YFLogxK
UR - http://www.scopus.com/inward/citedby.url?scp=84861808502&partnerID=8YFLogxK
U2 - 10.1007/978-1-61779-815-3_12
DO - 10.1007/978-1-61779-815-3_12
M3 - Article
C2 - 22610561
AN - SCOPUS:84861808502
SN - 1064-3745
VL - 879
SP - 179
EP - 193
JO - Methods in Molecular Biology
JF - Methods in Molecular Biology
ER -