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Data on the change in the protein profiles of human subcutaneous white adipocytes during differentiation and maturation

ABSTRACT: We analyzed changes in the protein profiles that occur during differentiation and maturation of cultured human subcutaneous white preadipocytes. We divided the three cell lines (Cell Line-1, Cell Line-2, and Cell Line-3) of Caucasian-derived subcutaneous preadipocytes into five stages: i) subcutaneous preadipocytes as stage-1, ii) after inducing differentiation into adipocytes as stage-2, iii-v) from the initiation of lipid droplet formation to become mature subcutaneous adipocytes was defined as stage-3 to stage-5, depending on lipid droplet amount and formation. Proteins from the cells were extracted at each stage (stage-1 to stage-5), proteolytically cleaved using trypsin, and analyzed using untargeted liquid chromatography and mass spectrometry.

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Aya Kitamoto

PROVIDER: PXD035060 | JPOST Repository | Mon Jul 04 00:00:00 BST 2022

REPOSITORIES: jPOST

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Updating "Data on changes in lipid profiles during the differentiation and maturation of human subcutaneous white adipocytes analyzed using chromatographic and bioinformatic tools" with data on the changes in protein profiles.

Kitamoto Takuya T Kitamoto Aya A

Data in brief 20230307

Previously, we reported changes in the lipid profile of cultured human subcutaneous white preadipocytes during their differentiation and maturation. Here, using the same cells, we report changes in the protein profiles during differentiation and maturation as multi-omics data. The three cell lines of Caucasian-derived subcutaneous preadipocytes were divided into five stages: stage-1, subcutaneous preadipocytes; stage-2, following induction of differentiation into adipocytes; stages-3 to -5, from ...[more]

PMID: 36950557

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Project description:To better understand the scale of gene expression changes that occur during the formation of mature adipocytes from preadipocytes, we compared and characterised the transcriptome profile of mesenchymal stromal cells derived from human adipose tissue, otherwise known as adipose-derived stromal cells (ASCs), undergoing adipocyte differentiation on day 1, 7, 14 and 21 (representing the early to late stage process of adipogenesis). Microarray technique was systematically employed to study gene expression in adipose-derived stromal cells during adipogenic differentiation over a 21 day period to identify genes that are important in driving adipogenesis in humans. We have undertaken an in-depth transcriptome analysis of adipogenesis in human adipose-derived stromal cells (ASCs) induced to differentiate into adipocytes in vitro. Genes were differentially expressed on days 1, 7, 14 and 21 post-induction and numbered 128, 218, 253 and 240 respectively. Up-regulated genes were associated with neural and blood vessel development, leukocyte migration, and tumor growth, invasion and metastasis. They also shared common pathways with certain obesity-related pathophysiological conditions. Down-regulated genes were associated with osteogenesis and the immune response. KLF15, LMO3, FOXO1 and ZBTB16 transcription factors were up-regulated throughout the differentiation process. CEBPA, PPARG, ZNF117, MLXIPL, MMP3 and RORB were up-regulated only on days 14 - 21, which coincides with the maturation of adipocytes and could possibly serve as candidates for controlling fat accumulation and the size of mature adipocytes. We identified genes that were up-regulated only on day 1 - 7 and day 14 - 21 that could serve as potential early and late-stage differentiation markers. This study reveals potential markers for the different stages in ASC adipogenic differentiation which could serve as good candidates to combat obesity and further link the biology of fat cell formation to the co-morbidities of obesity.

Project description:The utility of human pluripotent stem cells as a tool for understanding disease and as a renewable source of cells for transplantation therapies is dependent on efficient differentiation protocols that convert these cells into relevant adult cell types. Here we report the robust and efficient differentiation of human pluripotent stem cells into adipocytes. We found that inducible expression of PPARG2 in pluripotent stem cell-derived mesenchymal progenitor cells programmed their development towards an adipocyte cell fate. Using this approach, multiple human pluripotent cell lines were differentiated into adipocytes with efficiencies of 85% to 90%. These pluripotent stem cell-derived adipocytes retained their identity independent of transgene expression, could be maintained in culture for several weeks, expressed mature markers, and exhibited mature functional properties such as lipid catabolism in response to a beta-adrenergic stimulus. Global transcriptional and lipid metabolomic analyses further confirmed the identity and maturity of these pluripotent stem cell-derived adipocytes. Mesenchymal progenitor cells (MPCs) derived from human embryonic stem cells hESCs and induced pluripotent stem cells (iPSCs) along with adipose-derived stromal vascular cells (ADSVCs) were subjected to induction of PPAR2 and compared to primary fat samples. Overall 2 ADSVC (ADSVC 24 nd 49) lines, 1 hESC (HUES9) line and 1 iPSC (BJRiPS) line were differentiated into MPCs, PPAR2 programmed, and compared to untreated MPCs and primary fat samples from 2 individuals. Each condition is either represented in duplicate or triplicate on affymetrix HuGene-1_0-st arrays. MPCs derived from the hESC lines HUES2 and HUES8, ADSVCs, and BJRiPS were also run on a separate platform (HG-U133_Plus_2) with more GEO presence to facilitate analysis (34 samples, two platforms total). Supplementary file(s): GeneSymbol-collapsed data represent the final normalized data used for analyses in the manuscript.

			Action	DRS
	CellLine-1_Stage-1.raw	Raw
	CellLine-1_Stage-2.raw	Raw
	CellLine-1_Stage-3.raw	Raw
	CellLine-1_Stage-4.raw	Raw
	CellLine-1_Stage-5.raw	Raw