Mammary epithelial cell subpopulations from control and ovariectomized mice
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE20401 | GEO | 2010/04/10
SECONDARY ACCESSION(S): PRJNA129827
REPOSITORIES: GEO
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Project description:To study the effect of pregnancy on mouse mammary epithelial subpopulations, epthelial cells derived from control or ovariectomized adult mice were isolated using fluorescence-activated cell sorting. After elimination of haematopoietic and endothelial cells, two distinct epithelial subpopulations were sorted using antibodies against CD29 and CD24. Based on the immunohistochemical phenotype, and in vivo and in vitro functional assays, these subpopulations were identified as mammary stem cell enriched (CD29hiCD24+) and luminal (CD29loCD24+) respectively (ref: Shackleton et al, Nature 2006). Microarray profiling was used to compare gene expression profiles of the two subpopulations isolated from ovariectomized and control mice. For each of two biological replicates, 6 FVB/NJ mice were ovariectomized at 8 weeks of age. 4 weeks after ovariectomy (Ovx) mammary gland from control or Ovx animals were collected and digested to obtain a single cell suspension. CD45-CD31-TER119- cells were then sorted based on the expression of cell surface markers CD24 and CD29. Similarly for two pools of control mice. There were also 4 technical replicates, to make 12 BeadChips in total.
2010-04-10 | E-GEOD-20401 | biostudies-arrayexpress
Project description:To study the effect of pregnancy on mouse mammary epithelial subpopulations, epthelial cells derived from virgin or pregnant (12.5 day pregnant) mice were isolated using fluorescence-activated cell sorting. After elimination of haematopoietic and endothelial cells, two distinct epithelial subpopulations were sorted using antibodies against CD29 and CD24. Based on the immunohistochemical phenotype, and in vivo and in vitro functional assays, these subpopulations were identified as mammary stem cell enriched (CD29hiCD24+) and luminal (CD29loCD24+) respectively (ref: Shackleton et al, Nature 2006). Microarray profiling was used to compare gene expression profiles of the two subpopulations in 12.5 day pregnant and virgin mice.
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Project description:To study the effect of pregnancy on mouse mammary epithelial subpopulations, epthelial cells derived from virgin or pregnant (12.5 day pregnant) mice were isolated using fluorescence-activated cell sorting. After elimination of haematopoietic and endothelial cells, two distinct epithelial subpopulations were sorted using antibodies against CD29 and CD24. Based on the immunohistochemical phenotype, and in vivo and in vitro functional assays, these subpopulations were identified as mammary stem cell enriched (CD29hiCD24+) and luminal (CD29loCD24+) respectively (ref: Shackleton et al, Nature 2006). Microarray profiling was used to compare gene expression profiles of the two subpopulations in 12.5 day pregnant and virgin mice. For each biological replicate, mammary gland from virgin or 12.5 day pregnant FVB/NJ mice were collected and digested to obtain a single cell suspension. CD45-CD31-TER119- cells were then sorted based on the expression of cell surface markers CD24 and CD29. There were 4 pools of pregant mice and 3 pools of virgin mice.
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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.
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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.
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Project description:Using a syngeneic p53 null mouse mammary gland tumor model that closely mimics human breast cancer, we have identified by limiting dilution transplantation as well as in vitro mammosphere and clonogenic assays a Lin-CD29HighCD24High subpopulation of tumor-initiating cells. Differentially expressed genes in the Lin-CD29HighCD24High mouse mammary gland tumor-initiating cell population include those involved in DNA damage response and repair, as well as genes involved in epigenetic regulation previously shown to be critical for stem cell self-renewal. Experiment Overall Design: The cells were labeled (for FACS sorting) with CD29 and CD24 antibodies, and 4 subpopulations were collected (CD29HighCD24High, CD29HighCD24Low,CD29LowCD24High, and CD29LowCD24Low) to do either in vivo transplantation or to isolate RNA from each of them for array to correlate the in vivo data. Experiment Overall Design: 12 samples (RNA of 4 subpopulations based on expression of CD29 and CD24 for each tumor of 3) were included in the identification of differentially expressed genes of tumor-initiating cells. 5 samples (RNA of subpopulations based on expression of CD29 and CD24 of normal mammary epithelial cells ) were included in the normal group analysis. Reference RNA was used to normalize all samples.
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Project description:MaSC, Luminal progenitor 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), and the transcirptome profiles were determined and compared.
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Project description:MaSC and luminal 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), and the microRNA profiles were determined using microRNA array and compared.
2017-05-05 | GSE85634 | GEO
Project description:The Wnt/beta-catenin signalling pathway plays a central role in mammary stem cell homeostasis and in breast cancer. We employed the CD29hiCD24+ cell surface antigens to identify a subpopulation of mammary CSCs from Apc1572T/+, a mouse model for metaplastic breast adenocarcinoma, a subtype of triple-negative breast cancer in man. The MaCSCs are capable of recapitulating tumorigenesis when transplanted at low multiplicities in vivo, and of forming self-renewing organoids in vitro. Expression profiling of the different subpopulations sorted from normal and neoplastic mammary tissues revealed that the normal stem cell compartment is more similar to tumor cells than to their own differentiated progenies. Accordingly, Wnt signaling was found to be activated in the subpopulation encompassing normal mammary stem cells, though to a lesser degree than in the tumor cells. By comparing normal with cancer mouse mammary compartments, we were able to derive a MaCSC-specific signature composed of human orthologous genes able to predict poor survival, relapse and distant metastasis in human breast cancer. Finally, upon intravenous injection, only MaCSCs among the different tumor cell subpopulations are able to form metastatic lesions in a broad spectrum of anatomical sites. Overall, our data indicate that constitutive Wnt signaling activation interferes with mammary stem cell homeostasis leading to metaplasia and basal-like adenocarcinomas. The objective was to compare the: i) expression profiles between normal adult mammary stem cells and tumor cancer stem cells identified by Lin-CD29hiCD24+; ii) expression profile between adult stem cells and their differentiated counterparts both in normal and in tumor tissue to generate a cancer stem cell signature that can be used to compare with mammary human tumor expression data to predict survival and prognosis. To this aim five independent mammary adenocarcinomas from C57BL6/J Apc+/1572T mice and three independently isolated pools of mammary glands from C57BL6/J Apc+/+ mice were employed to sort 10,000 cells of each of the following populations: Lin-, Lin-CD29+CD24+ and Lin-CD29hiCD24+.
2013-08-29 | E-GEOD-40704 | biostudies-arrayexpress
Project description:The Wnt/beta-catenin signalling pathway plays a central role in mammary stem cell homeostasis and in breast cancer. We employed the CD29hiCD24+ cell surface antigens to identify a subpopulation of mammary CSCs from Apc1572T/+, a mouse model for metaplastic breast adenocarcinoma, a subtype of triple-negative breast cancer in man. The MaCSCs are capable of recapitulating tumorigenesis when transplanted at low multiplicities in vivo, and of forming self-renewing organoids in vitro. Expression profiling of the different subpopulations sorted from normal and neoplastic mammary tissues revealed that the normal stem cell compartment is more similar to tumor cells than to their own differentiated progenies. Accordingly, Wnt signaling was found to be activated in the subpopulation encompassing normal mammary stem cells, though to a lesser degree than in the tumor cells. By comparing normal with cancer mouse mammary compartments, we were able to derive a MaCSC-specific signature composed of human orthologous genes able to predict poor survival, relapse and distant metastasis in human breast cancer. Finally, upon intravenous injection, only MaCSCs among the different tumor cell subpopulations are able to form metastatic lesions in a broad spectrum of anatomical sites. Overall, our data indicate that constitutive Wnt signaling activation interferes with mammary stem cell homeostasis leading to metaplasia and basal-like adenocarcinomas. The objective was to compare the: i) expression profiles between normal adult mammary stem cells and tumor cancer stem cells identified by Lin-CD29hiCD24+; ii) expression profile between adult stem cells and their differentiated counterparts both in normal and in tumor tissue to generate a cancer stem cell signature that can be used to compare with mammary human tumor expression data to predict survival and prognosis. To this aim five independent mammary adenocarcinomas from C57BL6/J Apc+/1572T mice and three independently isolated pools of mammary glands from C57BL6/J Apc+/+ mice were employed to sort 10,000 cells of each of the following populations: Lin-, Lin-CD29+CD24+ and Lin-CD29hiCD24+.
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