Transcriptomics,Genomics

Dataset Information

29

Gene expression profiles of epithelial and mesenchymal cell populations extracted from normal human breast tissue


ABSTRACT: Stromal-epithelial interactions play a fundamental role in tissue homeostasis, controlling cell proliferation and differentiation. The goals and objectives of this study were 1.) To generate gene expression profiles and identify signatures of freshly isolated, non-cultured, luminal epithelial or basal epithelial and CD10 positive or negative stromal cell populations from the human breast. 2.) Use the generated signatures to characterize and validate the molecular identity of human primary epithelial and stromal/mesenchymal breast cells maintained long-term in novel ex vivo culture conditions in serum free medium (previous study in our lab) Breast tissue from three different reduction mammoplasties was dissociated and single cells were labeled with antibodies against lineage markers (CD31, CD45 CD235ab), CD10, EpCAM, CD49f and DAPI. Different cell populations were identified, separated and sorted with a fluorescence activated cell sorter. Gene expression profiling of luminal epithelial (EpCAM+, CD49f+/-), basal epithelial (EPCAM-, CD49f+), CD10 negative and CD10 positive stromal cells was performed and distinct gene expression signatures for the four cell populations were identified. Overall design: A total of 12 samples were analyzed. To obtain three complete biological replicates, breast tissue from three different reduction mammoplasties (MP9, MP13, MP15) was dissociated and single cells were labeled with fluorescent antibodies prior to cell sorting. Four distinct cell populations (two epithelial and two stromal) were identified and gated. 300 cells of each of the four cell populations were sorted directly in 100ul TRIZOL for all three donor tissues. The samples were immediately frozen on dry ice and stored at -70°C prior to RNA extraction. Gene expression profiles of luminal epithelial cells (EpCAM+, CD49f+/-), basal epithelial (EPCAM-, CD49f+), CD10 negative and CD10 positive stromal cells were successfully obtained for all three reduction mammoplasties.

INSTRUMENT(S): [HuGene-1_0-st] Affymetrix Human Gene 1.0 ST Array [transcript (gene) version]

SUBMITTER: Tim Roloff  

PROVIDER: GSE51200 | GEO | 2015-09-26

SECONDARY ACCESSION(S): PRJNA221582

REPOSITORIES: GEO

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