Unknown,Transcriptomics,Genomics,Proteomics

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Comparison of gene expression profiles between human DFAT cells and ADSCs derived from same donors

ABSTRACT: Dedifferentiated fat (DFAT) cells, established in vitro from mature adipocytes, exhibit properties of multipotent mesenchymal stem/stromal cells (MSCs), such as the ability to differentiate into multiple mesenchymal lineages. Although DFAT cells exhibit certain properties of proliferative progeny, at present there is only limited knowledge about their characteristics as MSCs because those cells are considered to be potential artifacts of cell culture. To elucidate the identity of DFAT cells, we compared gene expression profiles of human DFAT cells and adipose-derived stem/stromal cells (ADSCs) established using adipose tissue from the same donors. Microarray analysis showed that the global RNA expression profiles of human DFAT cells were very similar to those of ADSCs, a representative MSC, despite being committed adipocyte progenitors. Subcutaneous adipose tissues that were obtained during surgical operation for non-malignant disease were donated by 3 patients after obtaining informed consent. Three sets of DFAT cells and ADSCs, each derived from adipose tissue from the same donor were used for RNA extraction and subsequent microarray analysis.

ORGANISM(S): Homo sapiens

SUBMITTER: Yuji Iribe

PROVIDER: E-GEOD-47869 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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Project description:Adipogenesis occurs through a specific gene program in undifferentiated fat progenitors. We hypothesized that the properties of the fat progenitors are regulated by hox genes, the developmental genes essential in different tissue stem cells. Their biased expression in white and brown fat implies roles in distinguishing the two fat types. Among 39 Hox genes, Hoxc8 is highly enriched in undifferentiated adipose tissue stem cells (ADSCs) and down-regulated in differentiated adipocytes. Forced expression of Hoxc8 suppressed adipocyte differentiation of ADSCs. Using microarrays, we investigated the effect of Hoxc8 overexpression on global transcripts in ADSCs. We compared among four groups: untreated ADSCs, adipogenic induction media (MDI)-treated ADSCs, MDI-treated ADSC-vector and MDI-treated ADSC-Hoxc8. A number of, but not all, adipogenesis-related genes are suppressed by Hoxc8. This dataset illustrates the global effect of Hoxc8, a developmental transcription factor, on the expression of adipogenesis-related genes. Gene expression was compared among untreated ADSCs (control), adipogenic induction media-treated ADSCs, adipogenic induction media-treated ADSC-vector (ADSCs transduced with control vector), and adipogenic induction media-treated ADSC-Hoxc8 (ADSCs transduced with human Hoxc8). Total RNA was isolated from ADSCs using the Qiagen RNeasy kit (Qiagen). At NimbleGen, quality and yield were verified before cDNA synthesis and Cy3-end labeling. The labeled cDNA samples were hybridized to Homo sapiens 4-Plex arrays (Roche NimbleGen, A4487001-00-01) that represent 24,000 human genes. Raw data files for each sample were normalized and background-corrected using a Robust Multi-Array Analysis as implemented by NimbleScan software. Students’ two-tail t-tests were conducted among the samples for each transcript and fold-change was determined. Transcripts whose abundance was significantly altered (P < 0.05) and an absolute fold change greater than 2 were defined as differentially regulated.

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Comparison of gene expression profiles between human DFAT cells and ADSCs derived from same donors

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