Project description:Transcriptome response to IFNa treatment in MaSC, Luminal progenitor cells were studied. The MaSC, Luminal progenitor cells enriched subpopulations were sorted based on CD24/CD29 expression from mammary epithelial cells of virgin female mice . (Tiede BJ et al., 2009, PLoS ONE 4(11): e8035. doi:10.1371/journal.pone.0008035). The cells were cultured in mammosphere condition in vitro and treated with Ifna for 8 days. The total RNA samples are collected and compared using microarray analysis.
Project description:In this study we mapped H3K27me3, H3K4me3 and H3K9me2 marks in three mammary epithelial subsets: MaSC/basal (MS), luminal progenitor (LP) and mature luminal (ML) in the steady-state. In addition, profiles were generated for H3K4me3 and H3K27me3 marks in the MaSC/basal and luminal populations from the glands of ovariectomized or mid-pregnant (12.5 days) mice as well as from control virgin mice.
Project description:The purpose of this microarray experiment was to obtain reference gene expression patterns of a number of epithelial cell populations [mammary stem cells (MASC), luminal progenitors (LP), alveolar luminal stem/progenitor cells (WC virgin-these are mammary epithelial cells genetically marked by Wap-Cre in virgin females), mature luminal cells (ML, mainly represent ductal luminal cells in virgin females), and alveolar luminal cells (WC preg M-bM-^@M-^S these are alveolar cells genetically marked by Wap-Cre during mid-gestation)] present in the mammary gland of wildtype adult mice on a C57BL6 genetic background. For the isolation of RNA from mammary stem cells (MASC, Lin-CD24+CD29hi), luminal progenitors (LP, Lin-CD24hiCD29+CD61+), and mature luminal cells (ML, lin-CD24hiCD29+CD61-), the thoracic and inguinal mammary glands from 3 adult virgin female mice were harvested, minced and digested into a single cell suspension. Form each of these 3 single cell suspensions, the above populations were sorted by FACS. For alveolar luminal stem/progenitor cells and alveolar luminal cells, Lin-YFP+ mammary epithelial cells were isolated from virgin or midgestation mice genetically marked by Wap-Cre;R26Y. R26Y is a conditional YFP reporter that would be turned on upon Cre-mediated recombination.
Project description:To delineate epithelial subpopulations in mouse mammary tissue, hematopoietic and endothelial cells were depleted from freshly isolated cell suspensions derived from mammary glands using fluorescence-activated cell sorting. The resultant Lin- population was fractionated into four distinct subpopulations using antibodies against CD29, CD24 and CD61. Based on the immunohistochemical phenotype, and in vivo and in vitro functional assays, these subpopulations were identified as fibroblast-containing stromal (CD29loCD24-), mammary stem cell (MaSC)-enriched (CD29hiCD24+CD61+), luminal progenitor (CD29loCD24+CD61+), and mature luminal (CD29loCD24+CD61-) cell subpopulations. Microarray profiling was used to derive gene expression signatures of these 4 subpopulations. The four mammary cell subpopulations were found to have distinct gene expression profiles. Four mammary cell subpopulations from 3-5 pooled mouse tissues were analysed. MS is for the MaSC-enriched cell subpopulation. LP is for the Luminal Progenitor subpopulation. ML is for the Mature Luminal subpopulation.
Project description:In this study we mapped H3K27me3, H3K4me3 and H3K9me2 marks in three mammary epithelial subsets: MaSC/basal (MS), luminal progenitor (LP) and mature luminal (ML) in the steady-state. In addition, profiles were generated for H3K4me3 and H3K27me3 marks in the MaSC/basal and luminal populations from the glands of ovariectomized or mid-pregnant (12.5 days) mice as well as from control virgin mice. We used ChIPseq analysis to determine histone modification marks in the MS, LP and ML subsets in the steady-state. We then determined the histone modification profiles of MS and sorted luminal (Lum) cells from pregnant (12.5 dP) or ovariectomized (OVX) mice and compared these with the profiles of control virgin mice in order to study the effect of hormones on the mammary epigenomes.
Project description:MaSC, luminal progenitor enriched subpopulations and total MECs as well, were isolated from both wild type and ∆Np63 KO heterozygous mouse and the transcriptome profiles were determined and compared.
