Project description:Characterization of gene expression levels with RNA-seq was performed on self-renewing (SR) and senescent (SEN) human adult adipose derived mesenchymal stem cells (hADSCs) using the Roche 454 pyrosequencing platform.
Project description:Mouse spermatogonial stem cells (SSCs) continuously self-renew on the feeder layers in serum-free culture medium supplemented with glial cell line-derived neurotrophic factor and fibroblast growth factor 2. To identify novel nuclear proteins involved in SSC maintenance, comparative proteomic profiling of nuclear proteins was performed between self-renewing and differentiation-initiated SSCs in culture. The self-renewing SSC cultures were established from C57BL/6 mouse testes. Nuclear fractions from self-renewing SSC cultures treated with ethanol as a vehicle control (spermatogonial stem cells) and differentiation-initiated SSC cultures treated with 0.3 μM retinoic acid for 24 h (spermatogonial progenitor cells) were isolated for proteomic analysis.
Project description:In this study, we have addressed how cellular senescence influences the immunomodulatory potential of human mesenchymal stem cells (hMSCs). We induced cell senescence in a panel of bone marrow-derived hMSC samples by means of gamma-irradiation, and performed both gene expression and miRNA microarray analyses on the untreated and senescent samples. We also compared the gene expression profile of untreated and senescent hMSCs with those obtained from several hMSCs samples used in an ongoing allogeneic clinical study of Graft Versus Host Disease (GVHD), of which their therapeutic efficacy is known. We have identified several genes (PLEC, C8orf48, TRPC4, and ZNF14) differentially expressed in senescent hMSCs that are similarly regulated in hMSC samples that did not show a therapeutic effect in the GVHD study. These genes might be useful as markers to evaluate the therapeutic potential of hMSCs used in future clinical studies. We compared the gene and miRNA expression profiles of untreated (WT) control bone marrow-derived hMSCs with the same primary cell lines 10 days after gamma-irradiation (SEN) and with several hMSCs samples used in a clinical stydy of GVHD. The samples used in the clinical study were classified in two groups, depending on whether they elicited a therapeutic response (G1) or not (G2). A total of four independent samples (biological replicates) were used for WT and SEN conditions. For the samples used in the clinical study, a total of five samples were used for the G1 group, and three samples for the G2 group.
Project description:To understand the differentiation of ovarian cancer stem cells (CSCs), We derived two phenotypes of CSCs and identified the gene expression profiling. The CSCs were derived from Cp70 ovarian cancer cells and cultured in suspension and examined every day for sphere formation. Spheres were then dissociated and passaged at least eight times in 2 months to generate spheres, which are henceforth referred to as SR cells. The surface of SR-I was smooth regardless of the size, whereas SR-II was morula. SR-I could differentiate into multiple-lineage cell types under specific induction conditions. SR-I spheroids could differentiate to SR-II spheroids through epithelial-mesenchymal transition.The self-renewal was slower for SR-I than for SR-II.
Project description:The Wnt target gene Lgr5 marks actively dividing stem cells in Wnt-driven, self-renewing tissues such as small intestine and colon, stomach and hair follicles. A 3D culture system allows long-term clonal expansion of single Lgr5+ stem cells into transplantable organoids that retain many characteristics of the original epithelial architecture. A crucial component of the culture medium is the Wnt agonist Rspo, the recently discovered ligand of Lgr5. Here we show that Lgr5-LacZ is not expressed in healthy adult liver, yet that small Lgr5-LacZ+ cells appear near bile ducts upon damage, coinciding with robust activation of Wnt signaling. As shown by lineage tracing using a novel Lgr5-ires-CreERT2 knock-in allele, damage-induced Lgr5+ cells generate hepatocytes and bile ducts in vivo. Single Lgr5+ cells from damaged liver can be clonally expanded as organoids in Rspo1-based culture medium over multiple months. Such clonal organoids can be induced to differentiate in vitro and to generate functional hepatocytes upon transplantation into FAH-/- mice. These findings imply that previous findings on Lgr5+ stem cells in actively self-renewing tissues extend to damage-induced stem cells in a tissue with a low rate of spontaneous self-renewal. We first generated arrays from multiple wildtype tissues including muscle, white adipose tissues, brown adipose tissues, liver and pancreas. Then we generated arrays from liver derived cultures maintained in different conditions, and compared the expression profile with the corresponding parental tissues and other non-related tissues.
Project description:Adipose tissue harbours a significant number of multipotent adult stem cells of mesenchymal origin known as adipose-derived stem cells (ADSCs). Broad differentiation potential and convenient accessibility of ADSCs make them an attractive source of adult mesenchymal stem cell for regenerative medicine and cell developmental plasticity research. Genome-wide microarray expression profiling was performed to identify genes deregulated during osteogenic differentiation of ADSCs to evaluate developmental plasticity of these cells. Dynamics of epigenetic modifications were analyzed in parallel and associated with the gene expression profile. Gene expression profile was analyzed in adipose-derived stem cells (ADSCs) differentiated into osteogenic lineage from 3 donors and compared to undifferentiated cells from the same donors.
Project description:Tissue macrophages derive from bone marrow monocytes, and recent studies using mice have revealed that they also derive from yolk sac precursors or fetal liver monocytes. However, embryo-derived macrophages are supposed to be more important to maintain tissue macrophage pool because they can self-renew. Here, we show that adult bone marrow-derived macrophages (MDM) also retain the ability of self-renewal. Where they were readily obtained by a long-term culture: mouse bone marrow cells were cultured with macrophage colony-stimulating factor (M-CSF). After several passages, most MDM died owing to their limited life span with survival and expansion of self-renewing macrophages resided in a small fraction. Self-renewing macrophages were not tumorigenic, but proliferate for a long period in almost unlimited numbers. Despite being distinct from MDM, they were phenotypically and functionally differentiated macrophages, and could differentiate into dendritic cells or osteoclasts. Moreover, Krüppel-like Factor 2 (KLF2) involved in self-renewal of embryonic stem cells, was markedly up-regulated by M-CSF-stimulation in self-renewing macrophages, which was accompanied with a gradual down-regulation of MafB, a suppressor of KLF2 expression. Importantly, knockdown of KLF2 as well as c-Myc caused cell cycle arrest, apoptosis, and diminished cell growth. Our culture method results suggest the presence of precursor(s) for self-renewing macrophages in adult bone marrow that can be used to describe discrepancy of adult- and embryo-derived macrophages. Microarray data from both monocyte-derived and bone marrow-derived mouse macrophages are used to detail the global gene expression profile underlying phenotype, function and self-renewal capacity in order to unravel difference/similarity in phenotype/function and mechanism/degree of self-renewal between the two distinct macrophages.
Project description:Adipose tissue harbours a significant number of multipotent adult stem cells of mesenchymal origin known as adipose-derived stem cells (ADSCs). Broad differentiation potential and convenient accessibility of ADSCs make them an attractive source of adult mesenchymal stem cell for regenerative medicine and cell developmental plasticity research. Genome-wide microarray expression profiling was performed to identify genes deregulated during osteogenic differentiation of ADSCs to evaluate developmental plasticity of these cells. Dynamics of epigenetic modifications were analyzed in parallel and associated with the gene expression profile.
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