Dystrophic single myofibers vs. control single myofibers
Ontology highlight
ABSTRACT: MicroRNA-expression profile of dystrophic single fibers compared to wild type single fibers isolated from different muscles of mdx and C57BL mice. Myofibers were isolated from different muscle types (tibialis, diaphragm and quadriceps) of gender- (male) and age- (3 month old and half) matched wt and dystrophic mice. 9 total samples per animal model (C57BL, mdx), 3 replicates per muscle type.
Project description:While enucleation is a critical step in the terminal differentiation of human red blood cells, the molecular mechanisms underlying this unique process remain unclear. To investigate erythroblast enucleation, we studied the erythroid differentiation of human embryonic stem cells (hESCs), which provide a unique model for deeper understanding of the development and differentiation of multiple cell types. First, using a two-step protocol, we demonstrated that terminal erythroid differentiation from hESCs is directly dependent on the age of the embryoid bodies. Second, by choosing hESCs in two extreme conditions of erythroid culture, we obtained an original differentiation model which allows one to study the mechanisms underlying the enucleation of erythroid cells by analyzing the gene and miRNA (miR) expression profiles of cells from these two culture conditions. Third, using an integrated analysis of mRNA and miR expression profiles, we identified five miRs potentially involved in erythroblast enucleation. Finally, by selective knockdown of these five miRs we found miR-30a to be a regulator of erythroblast enucleation in hESCs. We compared the miRNA expression profles of sample groupe A (EB9) to sample goupe B (EB20). Microarray analysis was performed in triplicate for both sample groups at each time point: Day0, cells at the level of embryoid body (A-D0, B-D0) and Day18 of erythroid culture(A-D18, B-D18).
Project description:We showed that some microRNAs could be characteristic of the progression from adenoma to adenocarcinoma in colorectal cancer. 48 colorectal biopsy samples (28 adenomas, 15 adenocarcinomas and 5 normal mucosae) were analyzed. We generated three comparisons: adenomas versus. normal mucosae, adenocarcinomas versus. normal mucosae, and adenocarcinomas versus. adenomas.
Project description:This SuperSeries is composed of the following subset Series: GSE30064: Cultured human amniotic fluid-derived mesenchymal stromal cells [PIQOR data] GSE30065: Cultured human amniotic fluid-derived mesenchymal stromal cells [miRXplore data] Refer to individual Series
Project description:While enucleation is a critical step in the terminal differentiation of human red blood cells, the molecular mechanisms underlying this unique process remain unclear. To investigate erythroblast enucleation, we studied the erythroid differentiation of human embryonic stem cells (hESCs), which provide a unique model for deeper understanding of the development and differentiation of multiple cell types. First, using a two-step protocol, we demonstrated that terminal erythroid differentiation from hESCs is directly dependent on the age of the embryoid bodies. Second, by choosing hESCs in two extreme conditions of erythroid culture, we obtained an original differentiation model which allows one to study the mechanisms underlying the enucleation of erythroid cells by analyzing the gene and miRNA (miR) expression profiles of cells from these two culture conditions. Third, using an integrated analysis of mRNA and miR expression profiles, we identified five miRs potentially involved in erythroblast enucleation. Finally, by selective knockdown of these five miRs we found miR-30a to be a regulator of erythroblast enucleation in hESCs. Refer to individual Series
Project description:Membrane vesicles released by neoplastic cells into extracellular medium contain potential of carrying arrays of oncogenic molecules including proteins and microRNAs (miRNA). Extracellular (exosome-like) vesicles play a major role in cell-to-cell communication. Thus, the characterization of miRNAs of exosome-like vesicles is imperative in clarifying intercellular signaling as well as identifying disease markers. microarray analysis identified several oncogenic miRNA between the two types vesicles. Exosome-like vesicles were isolated using gradient centrifugation from MCF-7 and MDA-MB 231 cultures. Sample-derived miRNA expression was analyzed on Miltenyi's miRXploreM-bM-^DM-" platform. The RNA was isolated from two replicate samples of MDA-MB231 and MCF7 cell lines. The miRNAs extracted from all four samples were labeled with Hy5 and hybridized against Hy3-labeled Universal Reference miRNA (UR). Candidate miRNAs were identified. In this report, the re-ratios of MDA1 or MDA2 relative to a virtual pool of both MCF7 samples 1 and 2 were used for further analyses. 1- Reratio No. 7 MDA 1 vs MCF7 1+2 reratios of (MDA 1 vs UR) and (MCF7 1 vs UR + MCF7 2 vs UR) 7970041, (7970039 + 7970040) 2- Reratio No. 8 MDA 2 vs MCF7 1+2 reratios of (MDA 2 vs UR) and (MCF7 1 vs UR + MCF7 2 vs UR) 7970042, (7970039 + 7970040) Exosome-like vesicles were isolated using gradient centrifugation from MCF-7 and MDA-MB 231 cultures. Sample-derived miRNA expression was analyzed on Miltenyi's miRXploreM-bM-^DM-" platform. The RNA was isolated from two replicate samples of MDA-MB231 and MCF7 cell lines. The miRNAs extracted from all four samples were labeled with Hy5 and hybridized against Hy3-labeled Universal Reference miRNA (UR). Candidate miRNAs were identified. In this report, the re-ratios of MDA1 or MDA2 relative to a virtual pool of both MCF7 samples 1 and 2 were used for further analyses. 1- Reratio No. 7 MDA 1 vs MCF7 1+2 reratios of (MDA 1 vs UR) and (MCF7 1 vs UR + MCF7 2 vs UR) 7970041, (7970039 + 7970040) 2- Reratio No. 8 MDA 2 vs MCF7 1+2 reratios of (MDA 2 vs UR) and (MCF7 1 vs UR + MCF7 2 vs UR) 7970042, (7970039 + 7970040)
Project description:Human amniotic fluid-derived mesenchymal stromal cells (hAMSC) have become one of the main cell populations used in regenerative medicine and for the study of various clinical disorders. These cells have a great capacity for proliferation and differentiation, do not form teratomas when transplanted into animal models and their stemness seems to be between embryonic cells and adult mesenchymal cells. Prior to their use in cell therapy they must be cultured and expanded in vitro, but the effect this process has on their fitness, a determining factor for the success or failure of cell therapy, is unknown. We undertook a follow-up of gene and microRNAs (miRNAs) expression using microarray of a hAMSC population kept under in vitro culture conditions for the first 15 passages. Significant changes were noted in the expression of various mRNAs and miRNAs, particularly down-regulation of TP53, increased expression of hsa-miR-125a and up-regulation of CDKN2D. The variations in TP53 and hsa-miR-125a may act as an indicator of the stemness of the hAMSC, whereas CDKN2D may indicate the reduction in the proliferative capacity of these cells in a TP53-independent mechanism. The genes described in this study will help evaluate the fitness of hAMSC, thus guaranteeing their biological quality for use in regenerative medicine. miRNA patterns were studied in in vitro culture samples of human amniotic fluid-derived mesenchymal stromal cells at passages P1, P3, P5 and P10 with miRXplorer microarrays using a synthetic pool of miRNAs. Cell pellets of 5 x 10E5 cells were cryopreserved in liquid nitrogen and sent to Miltenyi Biotec for their analysis.
Project description:MicroRNA-expression profile of dystrophic single fibers compared to wild type single fibers isolated from different muscles of mdx and C57BL mice.
Project description:MicroRNA-expression profile of dystrophic single fibres vs wild type single fibers isolated from different muscle of mdx and c57bl mice. Myofibers were isolated from different muscle type (tibialis, diaphragm and quadriceps of gender- (male) and age- (3 month old and half) matched wt and dystrophic mice). 9 total samples per animal model, 3 replicates per muscle type sample
Project description:MicroRNAs (miRNAs) are small non-coding RNAs that associate with Argonaute 2 protein to regulate gene expression at the post-transcriptional level in the cytoplasm. However, recent studies have reported that some miRNAs localize to and function in other cellular compartments. Mitochondria harbour their own genetic system that may be a potential site for miRNA-mediated post-transcriptional regulation. We aimed at investigating whether nuclear-encoded miRNAs can localize to and function in human mitochondria. To enable identification of mitochondrial-enriched miRNAs, we profiled the mitochondrial and cytosolic RNA fractions from the same HeLa cells by miRNA microarray analysis. Mitochondria were purified using a combination of cell fractionation and immunoisolation, and assessed for the lack of protein and RNA contaminants. We found 57 miRNAs differentially expressed in HeLa mitochondria and cytosol. Of these 57, a signature of 13 nuclear-encoded miRNAs was reproducibly enriched in mitochondrial RNA and validated by RT-PCR for hsa-miR-494, hsa-miR-1275 and hsa-miR-1974. This study provides the first comprehensive view of the localization of RNA interference components to the mitochondria. Our data outline the molecular bases for a novel layer of crosstalk between nucleus and mitochondria through a specific subset of human miRNAs that we termed ‘mitomiRs’. To assess whether nuclear-encoded miRNA are detectable in human mitochondria, we performed the following four steps approach. First, cultured HeLa cells were allowed to reach 80-100% confluence and subjected to fractionation in order to isolate the cytosolic fraction. From the same HeLa cells, mitochondria were isolated by immunomagnetic Anti-TOM22 MicroBeads from the Mitochondria Isolation Kit (Miltenyi Biotec). In total, six mitochondria preparations were perfromed, three of these were additionally treated with RNase A. Second, total RNA was extracted from the mitochondrial and cytosolic fractions. Third, mitochondrial and cytosolic RNA were respectively profiled by microRNA microarray analysis. Last, data were analyzed and normalized.Three independent assays were performed.
Project description:The goal of the current study was to identify miRNAs regulated by TGF-b in human CD8+ T cells and analyze the function of these miRNAs in shaping the immunomodulatory effect of TGF-b in these cells. We identified the miR-23a cluster to be upregulated and found that this cluster could target key molecules (IFN-g and LAMP1) involved in immune response by CD8+ T cells. CD8+ T cells were isolated and purified from healthy human peripheral blood of 5 donors and were activated using beads coupled to anti-CD2, anti-CD3 and anti-CD24. They were then treated with 5ng/ml TGF-b, with 1M-BM-5M SD-208 or left untreated. RNA from these cells were then isolated and used for generating miRNA microarrays.