Project description:Human adipose-derived mesenchymal stem cells were cultured either in hypoxia (Hx; <0.1% oxygen) or standard culture conditions (normoxia, Nx) over 48 hours in serum-free medium. Human tympanic membrane keratinocytes were cultured in standard culture conditions over 48 hours in serum-free medium.
Project description:This SuperSeries is composed of the following subset Series: GSE25068: PcG/TrxG profiling of differentially aged adipose-derived mesenchymal stem cells GSE25069: Whole-genome microarray of long-term cultured adipose derived mesenchymal stem cells from differentially-aged mice GSE25679: microRNA profiling of mesenchymal stem cells from adipose tissue of differentially aged mice Refer to individual Series
Project description:This project is a report of protemoic data of secretome from human adipose tissue-derived stem cells. The ADSCs were cultured in serum-free condition, and LC-MS/MS identification was conducted to analyze the ingredients in the secretome.
Project description:Adipose tissue is a plentiful and easily accessible source of mesenchymal stem cells that have been shown to be multi potent and possibly have immuno suppressive capacity(Zuk, Zhu et al. 2002; McIntosh, Zvonic et al. 2006; Prichard, Reichert et al. 2008). Several studies have identified hypoxia and the molecular effector of reduced oxygen tension, HIF1α to be essential in the differentiation processes, stem cell proliferation and survival as well as in maintenance of stem cell characteristics (Lennon, Edmison et al. 2001; Goda, Ryan et al. 2003; Wang, Fermor et al. 2005; Lin, Lee et al. 2006; Malladi, Xu et al. 2007; Ohnishi, Yasuda et al. 2007). The fact that oxygen has a central role in cell development and metabolism makes the regulation of oxygen tension a valuable tool in the attempt of exploiting stem cells to their full potential. In the current study we investigate the transcriptional changes caused by two weeks of monolayer hypoxic expansion of human adipose tissue derived stem cells (ASCs). Application of the XVivo hypoxic workstation (BioSpherix, Redfield, NY) made it possible to conduct the expansion of six ASC lines at four different hypoxic conditions in parallel covering the range from mild (15% oxygen) to full hypoxic (1% oxygen) conditions. In this comprehensive investigation the Illumina bead microarray platform was employed to give a profound and new insight in the molecular changes effectuated by hypoxic expansion. The findings of this study demonstrate the significance of biologic variation in the transcriptional response to mild hypoxic expansion and display evidence that full hypoxic growth conditions at 1% oxygen tension selects for a more prolific cell population of reduced chondrogenic potential. Keywords: hypoxia response To investigate the transcriptional response to hypoxia, adipose tissue derived stem cells (ASCs) from six female individuals were cultured for 14 days at different oxygen tensions (ambient, 15%, 10%, 5% and 1%). Gene expression levels at each oxygen tension were compared to ambient conditions (Ambient oxygen tension) using Illumina Human-6 v2.0 microarrays.
Project description:Human adipose tissue derived stem cells were differentiated to adipocytes in vitro. At the end of differentiation, cells were treated with siRNA targeting CD248 followed by exposure to 1% oxygen levels. Microarray analysis were performed to identify differentially regulated genes.
Project description:Human mesenchymal stem cells (MSC) derived from perirenal adipose tissue (PV) of living kidney donors were cultured under various conditions, namely (1) control (medium+foetal bovine serum(FBS)) or (2) control (medium+heat-inactivated FBS); (3) with mixed-lympohocyte reactions (MLR) in transwell culture systems for 4 days; (4) with mixed-lympohocyte reactions (MLR) in transwell culture systems for 7 days; or (5)with pro-inflammatory cytokines(IFNgamma, TNFalpha and interleukin 6). We used microarrays to detail the effect of the various culture conditions on MSC.
Project description:Mesenchymal stem cells (MSCs) have been shown to exert therapeutic effects on various autoimmune diseases. However, such therapeutic effect is not always achieved. Among many reasons, MSC culture methodologies may account for the these differences. It is known that oxygen concentration could profoundly affect the properties of MSCs. Therefore, we compared human umbilical cord derived MSCs cultured under hypoxic and normoxic conditions. We used microarrays to profile the gene expression of human umbilical cord derived MSCs under 10% oxygen and normoxia conditions.
Project description:Scope: Adipose tissue is regarded as a true endocrine organ. Recent studies showed that adipose tissue derived exosomes could serve as carrier of circulating miRNAs to regulate distant targets. However, the characteristics of exosomal proteins released from adipose tissue have not been investigated yet. Methods and Results: In this study, we conducted a complementary protein profiling on exosome-like vesicles derived from adipose tissue (ELV-AT) with Label-free Quantitative Proteomic Analysis. A total of 3229 ELV-AT proteins were identified, among which only 266 proteins have been annotated as adipokines. 3 undefined adipokine candidates (NPM3, STEAP3, and DAD1) were selected for further validation. These 3 proteins were expressed in both white and brown adipose tissue, upregulated during adipogenic differentiation of both 3T3-L1 cells and adipose derived stem cells (ADSCs). Expressions of NPM3, DAD1 in ELV-AT were significantly decreased in obese subjects compared with lean controls while obesity could not alter the expression of STEAP3. Conclusions: Our profiling study of the ELV-AT proteins expands the list of adipokines and highlights the pivotal role of exosomal adipokines in the regulation of multiple biological processes within adipose tissue