Project description:Purpose: To analyze cardiac fibroblasts gene expression at different time following induction of osteogenic differentiation Methods: Freshly isolated cardiac fibroblasts (Passage 0 or passage 1) were plated at a density of 2.5 x 104 cells/cm2 in growth medium. After overnight incubation, osteogenesis was induced using differentiation medium (α-MEM supplemented with 10% FBS, 10 nM dexamethasone (Sigma, D4902), 20 mM β-glycerol phosphate (Sigma, G9422), and 50 μM L-ascorbic acid (Sigma, A4403). Cardiac fibroblasts harvested at Day0 (before differentiation medium treatment), Day7, Day14 and Day21 were used for RNA sequencing. Results: Cardiac fibroblasts harvested at different time points following induction of differentiation revealed clusters of genes whose expression was significantly altered in a temporal specific manner. Genes regulating cell cycle that were highly expressed in undifferentiated cardiac fibroblasts were down-regulated at the onset of differentiation and remained at low expression levels throughout the duration of osteogenic differentiation, consistent with the principle that induction of differentiation is associated with reduced rates of proliferation. In contrast, genes that were minimally expressed in cardiac fibroblasts were induced in a specific temporal manner during the course of osteogenic differentiation and included sets of genes known to regulate inflammation, extracellular matrix proteins and cell metabolism. Conclusions: Cardiac fibroblasts subjected to osteogenic differentiation progressively adopted an osteogenic signature.
Project description:Purpose: To investigate the function of EphrinB2 in the regulation of osteogenic differentiation of cardiac fibroblasts, we established EphrinB2-overexpressing cardiac fibroblasts and corresponding control group by lentivirus.Methods:Adult cardiac fibroblasts were isolated from 6 to 8-week-aged C57BL/6 mice, passage 1 cardiac fibroblasts plated at a density of 2.5 x 104 cells/cm2 were infected with the EphrinB2-overexpressing or vector lentivirus at an MOI of 20 for 12h, after 48 hours’ infection, osteogenesis was induced using differentiation medium (α-MEM supplied with 10% FBS, 10 mM β-glycerophosphate, 100nM dexamethasone and 50 µg/mL L-ascorbic acid). Cardiac fibroblasts harvested at Day7 were used for RNA sequencing.
Project description:Comprehensive analyses of miRNAs expression were performed using miRNA microarrays during osteogenic differentiation of PDGFRa+ mesenchymal progenitors isolated from human skeletal muscle to identify miRNAs that are involved in osteogenesis of PDGFRa+ mesenchymal progenitors. PDGFRa+ mesenchymal progenitors were isolated from the gluteus medius muscles of two different patients, and then subjected to osteogenic induction. Expression of miRNAs was examined using miRNA microarrays at the time points of one week and two weeks after osteogenic induction and were compared with that of uninduced cells.
Project description:miRNA pattern of cardic myocytes, cardiac fibroblasts and cardiac endothelial cells isolated from adult mice hearts at different time points after transaortic constriction (TAC) or sham surgery.
Project description:Here we report the reprogramming of human fibroblasts to produce chemically-induced osteogenic cells (ciOG), and explore the potential uses of ciOG in bone repair and disease treatment. A chemical cocktail of RepSox, forskolin, and phenamil was used for osteogenic induction of fibroblasts by activation of RUNX2 expression. Following a maturation, the cells differentiated toward an osteoblast phenotype that produced mineralized tissue. Single-cell RNA sequencing analysis of ciOG identified unique clusters of cells, among which the cluster of active cells related to osteogenic cells was ~49% of the cells, or ~80% of active cells; another cluster of active cells (~12% of the cells) was related to fibroblasts. The reprogramming trajectory over time revealed the transition from fibroblasts to mainly osteogenic cells or to an inactive state.
Project description:In this study, the gene expression profiling and genome-wide accessible chromatin mapping of mouse cardiac fibroblasts isolated from the uninjured myocardium and the infarct at multiple time points corresponding to different differentiation states were performed by RNA sequencing (RNA-seq) and the assay for transposase-accessible chromatin with high-throughput sequencing (ATAC-seq), respectively.
Project description:Pluripotent P19CL6 embryonic carcinoma cells can be differentiated to a cardiac lineage by culture in the presence of DMSO. The goal of this study was to characterize temporal gene expression patterns associated with cardiogenic differentiation. Gene expression analysis was conducted on differentiating P19CL6 cells at several time points following induction with 1% DMSO. Samples were processed for analysis by Affymetrix GeneChip.
Project description:RNA sequencing on EphrinB2 overexpression cardiac fibroblasts and vector cardiac fibroblasts within or without induction of osteogenic differentiation in the 7th day.
Project description:Ability to perform osteogenic differentiation is one of the minimal criteria of mesenchymal stem cells (MSCs). Still, it is generally unknown whether osteogenic differentiation is universal cell fate or various phenotypically similar cell states. Besides this, MSCs and their secretomes are actively using for cell/cell-free therapy development, but systemic inter-source variation in MSCs secretomes, proteomes and differentiation mechanisms are still poorly understood. Therefore, here we compared proteomic and secretomic profiles of human mesenchymal cells from six sources: osteoblasts (bone), WJ-MSCs (Warton’s jelly), AD-MSCs (adipose), PDLSCs (tooth: Periodontal Ligament Stem Cells), DPSCs (tooth: Dental Pulp Stem Cells) and GFs (tooth: Gingival Fibroblasts). For experiments we used cells in early passages (3-5) isolated from 3-6 individuals. All cells were compared in standard cultivation and in the 10th day after induction of osteogenic differentiation.
Project description:To examine the impact of the NFIB/MLL1 complex on the transcriptome of C3H10T1/2 mesenchymal stem cells (MSCs) and to identify NFIB/MLL1-dependent genes during osteogenic differentiation, we performed RNA-sequencing (RNA-seq) analysis of six different samples at two time points.
