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Gene expression of canine mammary cancer stem-like cells co-cultured with tumor associated macrophages

ABSTRACT: The aim of our study was to evaluate gene expression of canine mammary cancer stem-like cells co-cultured with tumor associated macrophages. Two canine mammary cancer cell lines (CMT-U27 and P114) were stained using anti-Sca1 (stem cell antigen 1), anti-EpCAM (Epithelial cell adhesion molecule) and anti-CD44 antibody. The FACS analysis showed 0,02-0,05% of Sca1+/EpCAM+/CD44+ in each of the cell line. Cancer stem-like cells were collected using FACS Aria II then co-cultured with tumor associated macrophages and used for further analysis of gene expression ( using Agilent Gene Expression Hybridization Kit ). Canine mammary cancer cell lines were stained using anti-Sca1 (stem cell antigen 1), anti-EpCAM (Epithelial cell adhesion molecule) and anti-CD44 antibodies. Next using FACS Aria II and Sca1+/EpCAM+/CD44+ cells were collected and co-cultured with tumor associated macrophages. Then, total RNA was isolated and hybridized at Gene Expression microarray.

ORGANISM(S): Canis lupus familiaris

SUBMITTER: Magdalena KrÃ³l

PROVIDER: E-GEOD-73230 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Gene expression of canine mammary cancer stem-like cells co-cultured with tumor associated macrophages

Project description:The aim of our study was to evaluate gene expression of canine mammary cancer stem-like cells co-cultured with tumor associated macrophages. Two canine mammary cancer cell lines (CMT-U27 and P114) were stained using anti-Sca1 (stem cell antigen 1), anti-EpCAM (Epithelial cell adhesion molecule) and anti-CD44 antibody. The FACS analysis showed 0,02-0,05% of Sca1+/EpCAM+/CD44+ in each of the cell line. Cancer stem-like cells were collected using FACS Aria II then co-cultured with tumor associated macrophages and used for further analysis of gene expression ( using Agilent Gene Expression Hybridization Kit ).

2016-06-01 | GSE73230 | GEO

miRNA expression of canine mammary cancer stem cells

Project description:The aim of our study was to assess miRNA expression of canine mammary cancer stem cells. Three canine mammary cancer cell lines (CMT-U27, CMT-U309 and P114) were stained using Anti-Sca1 (Stem cell antigen 1) antibodies. The FACS analysis showed 0,3-1% of Sca1+ cells in each of the cell line. The cells were sorted (using FACS Aria II) as Sca1+ and Sca1- and subjected to further analysis of miRNA expression (using Agilent custom miRNA microarray).

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2023-12-08 | E-MTAB-6031 | biostudies-arrayexpress

Evaluation of gene expression between CD44+ and CD44- cells in colonospheres

Project description:To identify genes that are differentially expressed between CD44+ and CD44- cells in colonospheres, we have employed whole genome microarray expression profiling as a discovery platform. Colonospheres from four colon cancer patients were sorted into CD44+ and CD44- cells using the FACS Aria II Cell Sorter, and total RNA extracted from the cells were labeled with Cy3 and used for microarray analyses with Agilent Whole Human Genome Oligo Microarrays.

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Evaluation of gene expression between CD44+ and CD44- cells in colonospheres

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3’RNA-seq of post-differentiation murine CD142− and CD142+ ASPCs sorted with four different anti-CD142 antibodies

Project description:To characterize CD142+ ASPCs (Aregs) after exposure to an adipogenic cocktail we performed bulk RNA-seq (using BRB-seq) of total, CD142− and CD142+ mouse adipose stem and progenitor cells (ASPCs), sorted using four different anti-CD142 antibodies. ASPCs were collected as Lin− (CD31− CD45− TER119−) CD29+ CD34+ SCA1+ cells of the mouse subcutaneous stromal vascular fraction using FACS.

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3’RNA-seq of murine CD142− and CD142+ ASPCs freshly sorted with four different anti-CD142 antibodies

Project description:To validate that the robustness of Aregs' (CD142+ ASPCs') molecular identity regardless of the antibody used, we performed transcriptomic profiling of freshly isolated CD142− and CD142+ mouse adipose stem and progenitor cells (ASPCs) sorted using four different anti-CD142 antibodies. ASPCs were collected as Lin− (CD31− CD45− TER119−) CD29+ CD34+ SCA1+ CD142+/− cells of the mouse subcutaneous stromal vascular fraction using FACS.

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Defining the molecular properties of the repairing epithelium during DSS induced colitis

Project description:In order to understand the molecular changes associated with the establishment of the repairing epithelium following dextran sulfate sodium (DSS) induced injury of the colon we have isolated Epcam+Sca1+ expressing epithelial cells from repairing tissue and Epcam+ homeostatic intestine. Cells isolated by flow cytometry were subjected to RNA extraction and analysed by expression analysis

2017-11-30 | E-MTAB-5249 | biostudies-arrayexpress

HDAC inhibitors promote the pro-myogenic activity of Sca1-positive interstitial cells in regenerating dystrophic muscles

Project description:We report here on the identification and functional characterization of Sca1-positive muscle interstitial cells that contribute to regeneration or fibroadipogenic degeneration of dystrophic muscles and mediate the beneficial effects of HDAC inhibitors (HDACi) in mdx mice. We found that the phenotype adopted by these cells and their biological activity are influenced by changes in muscle environment. While Sca1-positive muscle interstitial cells from healthy muscles spontaneously adopt a fibro-adipogenic phenotype, Sca1-positive muscle interstitial cells isolated from dystrophic muscles of young mdx mice show a latent myogenic phenotype that is implemented by the exposure to HDACi. Co-culture assays in vitro and co-transplantation experiments in vivo demonstrate that HDACi also improve Sca1-positive muscle interstitial cell ability to enhance the differentiation potential of adjacent satellite cells. Importantly, HDACi-induced myogenic phenotype and pro-regeneration activity were not observed in Sca1-positive muscle interstitial cells isolated from muscles of old mdx mice. The different phenotype of Sca1-positive muscle interstitial cells from mdx mice at different stages of disease progression correlated with the stage-dependent beneficial effect of HDACi, which were effective only at early stages of disease. Transplantation of Sca1-positive muscle interstitial cells isolated from regenerating young muscles into muscles of old mdx mice restored HDACi ability to increase myofiber size. These results indicate that Sca1-positive muscle interstitial cells are key cellular determinants of disease progression and mediate the beneficial effect of HDACi in a mouse model of muscular dystrophy. Isolation of Sca1+ muscle interstitial cells from TSA treated DMD/MDX mice and MuSc from DMD/MDX mice

2012-03-31 | E-GEOD-34843 | biostudies-arrayexpress

Gene expression changes during hematopoietic cell differentiation

Project description:Gene expression analyses of hematopoietic stem cells (HSCs), progenitor cells (HPCs), and differentiated cell. Gene expressions of long-term HSCs (CD34-ckit+Sca1+Lineage-), short term HSCs (CD34+ckit+Sca1+Lineage-), Progenitor cells (ckit+Sca1- Lineage-), and differentiated cels (Lineage+) were examined by microarray. Results provide insight into the mechanism of hematopoietic cell differentiation. Long-term HSCs (CD34-ckit+Sca1+Lineage-), Short term-HSCs (CD34+ckit+Sca1+Lineage-), Progenitor cells (ckit+Sca1- Lineage-, and Lineage+), and differentiated cell (Lineage+) were sorted from mouse bone marraw and were examined by microarray. Results provide insight into the mechanism of hematopoietic cell differentiation.

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