Adult human fibroblasts grown in low oxygen and with FGF2 supplementation have the capacity to tip the healing outcome of skeletal muscle injury - by favoring regeneration response in vivo over scar formation. Here, we compare the transcriptomes of control adult human dermal fibroblasts and induced regeneration-competent (iRC) fibroblasts to identify transcriptional changes that may be related to their regeneration competence.We identified a unique gene-expression profile that characterizes FGF2 ...[more]
Project description:The current study set to determine the effects of Fibroblast Growth Factor 2 and cell culture surface (tissue culture plastic and glass) on the transcriptome of adult human dermal fibroblasts. Transcriptional profiles of adult human dermal fibroblasts grown in four experimental conditions (glass, glass and addition of FGF2, plastic, plastic and addition of FGF2) were compared. Comparison of the transcriptomes will allow to identify significantly differentially expressed genes due to FGF2 and surface treatment, which in turn will allow for identification of the pathways affected by these factors in the human adult fibroblasts. Transcriptome analysis of adult human dermal fibroblasts grown on tissue culture plastic and glass, with and without 4ng/ml FGF2, was performed in two biological replicates and two technical replicates for each treatment condition.
Project description:Adult human dermal fibroblasts reside in vivo under low oxygen tension. Thus, low oxygen culture conditions represent a physiological state for adult human dermal fibroblasts. We have also previously shown that low oxygen and addition of basic fibroblast growth factor (FGF2) lead to prolonged life-span of adult human dermal fibroblasts. Therefore, we set to determine effects of low oxygen and FGF2 on the gene expression signature of adult human dermal fibroblasts. This global analysis will allow identification of genes affected and pathways regulated by low oxygen and FGF2. Overall design: Adult human dermal fibroblasts were grown in four different experimental conditions (2% oxygen, 2% oxygen with addition of 4 ng/ml FGF2, 19% oxygen, and 19% oxygen with addition of 4 ng/ml FGF2) and their transcriptomes were compared. Current study includes one biological replicate and three technical replicates of each sample (total of 12 samples).
Project description:Comparison of gene expression profile of different types of cells: hESCs (H1), human adult dermal fibroblasts, cells from adult human ovarian cell culture, FACS sorted SSEA-4 positive cells from adult human ovarian culture and cells from human ovarian cancer cell culture.
Project description:Transcriptome analysis of of control inhibitor and miR200b inhibitor transfected Human Dermal Adult Fibroblasts (HDAF) compared with Human Dermal Microvascular Endothelial Cells (HMEC). Injury induced inhibition of miR200b induces angiogenesis at the wound edges which help in the healing process. We have characterised the effect of miR200b suppression in Human Adult Dermal Fibroblasts converts to endothelial cells through transcriptional profiling. In this dataset, we include the expression data obtained from control inhibitor and miR200b inhibitor transfected Human Dermal Adult Fibroblasts, as well as Human Dermal Microvascular Endothelial Cells (HMEC) as positive control. Overall design: 9 total samples were analyzed by using the Affymetrix GeneChip® Human Transcriptome Array 2.0
Project description:Comparison of whole genome gene expression profiles of human testis derived ES-like cells with pluripotent stem cells (human embryonic stem cell lines), adult human bone marrow derived mesenchymal stem cells and human dermal fibroblasts. Overall design: Microarray study with total RNA of three testis derived ES-like cells clusters of one indivual (1) cultured in three different culture conditions A, B,C and embryoid bodies (EB) derived from them at 4, 7, 10, 14 and 18 days of suspension culture. Biological duplicates of human ES cell lines (HUES-1, GFP-hES-3),human bone marrow derived MSC (BMMSC1 and 2) and human dermal fibroblasts (DFB1 and 2) (LONZA CC-2511) at different passages were used as comparison groups.
Project description:Comparison of whole genome gene expression profiles of human testis derived ES-like cells with pluripotent stem cells (human embryonic stem cell lines), adult human bone marrow derived mesenchymal stem cells and human dermal fibroblasts. Microarray study with total RNA of three testis derived ES-like cells clusters of one indivual (1) cultured in three different culture conditions A, B,C and embryoid bodies (EB) derived from them at 4, 7, 10, 14 and 18 days of suspension culture. Biological duplicates of human ES cell lines (HUES-1, GFP-hES-3),human bone marrow derived MSC (BMMSC1 and 2) and human dermal fibroblasts (DFB1 and 2) (LONZA CC-2511) at different passages were used as comparison groups.
Project description:PKC-δ inhibition with the selective inhibitor, rottlerin, resulted in potent downregulation of type I collagen expression and production in normal human dermal fibroblasts and abrogated the exaggerated type I collagen production and expression in fibroblasts cultured from affected skin from patients with the fibrosing disorder, systemic sclerosis (SSc). To elucidate the mechanisms involved in the ability of PKC-δ to regulate collagen production in fibroblasts, we examined the effects of PKC-δ inhibition on the transcriptome of normal and SSc-derived human dermal fibroblasts. We used microarrays to detail the effects of PKC-δ inhibition with rottlerin on the transcriptome of normal and SSc-derived human dermal fibroblasts and to identify gene networks that might be involved in the regulation of extracellular matrix by PKC-δ. Overall design: Early passage normal and SSc human dermal fibroblasts were incubated with rottlerin (50 µg/mL) for 24 hours, followed by RNA extraction of treated and untreated samples and hybridization on Affymetrix microarrays. Early passage fibroblasts derived from normal and SSc dermal biopsies were utilized in this study to ensure that phenotypes remained as close as possible to original biopsy tissue. SSc patient sample biopsies were isolated from leading edge of forearm lesion; normal biopsies were obtained from forearm dermis.
Project description:We describe a so far uncharacterized, embryonic and self-renewing Neural Plate Border Stem Cell (NBSC) population with the capacity to differentiate into central nervous and neural crest lineages. NBSCs can be obtained by neural transcription factor-mediated reprogramming (BRN2, SOX2, KLF4, and ZIC3) of human adult dermal fibroblasts and peripheral blood cells (induced Neural Plate Border Stem Cells, iNBSCs) or by directed differentiation from human induced pluripotent stem cells (NBSCs). Moreover, human (i)NBSCs share molecular and functional features with an endogenous NBSC population isolated from neural folds of E8.5 mouse embryos. Upon differentiation, iNBSCs give rise to either (1) radial glia-type stem cells, dopaminergic and serotonergic neurons, motoneurons, astrocytes, and oligodendrocytes or (2) cells from the neural crest lineage. Here we provide array-based methylation data of iNBSCs reprogrammed from adult dermal fibroblasts (ADF), iPSC-derived NBSCs and adult dermal fibroblasts. The data provided demonstrate robust changes in the methylation landscape after reprogramming of human adult dermal fibroblasts into iNBSCs.
Project description:The goal of this study was to determine the similarity between human dermal microvascular endothelial cells, induced endothelial cells from fibroblasts, and fibroblasts through RNA-seq expression analysis. RNA samples from independently induced cultures, plus fibroblast and human dermal microvascular endothelial cultures were converted into individual cDNA libraries using Illumina TruSeq methods and subjected to single-end 50 base-sequence analysis at 20-30 million read depths. Examination of one fibroblast culture, one human dermal mibrovascular endothelial cell culture, and two induced endothelial cell cultures.
Project description:Fibroblasts synthesize the extracellular matrix of connective tissue and play an essential role in maintaining tissue integrity. We have previously shown that mouse skin connective tissue, the dermis, is comprised of functionally distinct fibroblast lineages. However, the extent of fibroblast heterogeneity in human skin is unknown. Here, using a combination of spatial transcriptional profiling of human and mouse dermis and single cell transcriptional profiling of human dermal fibroblasts, we show that there are at least four distinct fibroblast populations in adult human skin. We define markers permitting prospective isolation of these cells and show that although marker expression is rapidly lost in culture, different fibroblast subpopulations retain distinct functionality in terms of Wnt signalling, T cell communication and the ability to support human epidermal reconstitution in organotypic culture. Furthermore, while some fibroblast subpopulations are spatially segregated, others are not. These findings have profound implications for normal wound healing and diseases characterized by excessive fibrosis, and suggest that ex vivo expansion or in vivo ablation of specific fibroblast subpopulations may have therapeutic applications. Overall design: Spatial RNA sequencing of human papillary versus reticular dermis for 3 individuals, and single cell RNA sequencing of dermal fibroblasts for a single individual.