MiRNAs regulated by Gadd45a during somatic cell reprogramming
ABSTRACT: Gadd45a can enhance somatic cell reprogramming significantly. To explore the roles of Gadd45a playing in reprogramming, we performed miRNA microarray to identify miRNAs and signals pathways that regulated by Gadd45a. miRNAs expression of MEFs was measured at day8 in reprogramming. Four samples were set: MEFs infected with SKO plus Flag, MEFs infected with SKO plus Gadd45a, MEFs infected with SKOM plus Flag and MEFs infected with SKOM+Ga.
Project description:Gadd45a can enhance somatic cell reprogramming significantly. To explore the roles of Gadd45a playing in reprogramming, we performed whole genome microarray to identify genes and signals pathways that regulated by Gadd45a. Genes expression of MEFs was measured at day8 in reprogramming. Three samples were set: MEFs infected with SKO plus Flag, MEFs infected with SKO plus Gadd45a and MEFs infected with SKO plus G39A which is a negative mutant of Gadd45a.
Project description:Gadd45a can enhance somatic cell reprogramming significantly. To explore the roles of Gadd45a playing in reprogramming, we performed miRNA microarray to identify miRNAs and signals pathways that regulated by Gadd45a. Overall design: miRNAs expression of MEFs was measured at day8 in reprogramming. Four samples were set: MEFs infected with SKO plus Flag, MEFs infected with SKO plus Gadd45a, MEFs infected with SKOM plus Flag and MEFs infected with SKOM+Ga.
Project description:The direct conversion, or trans-differentiation, of non-cardiac cells into cardiomyocytes by forced expression of transcription factors and microRNAs provide promising ways of cardiac regeneration. However, genetic manipulations are still not desirable in real clinical applications. we report the generation of automatically beating cardiomyocyte-like cells from mouse fibroblasts with only chemical cocktails. These chemical-induced cardiomyocyte-like cells (CiCMs) express cardiomyocyte-specific markers, exhibit sarcomeric organization, and possess typical cardiac calcium flux and electrophysiological features. Microarray-bassed gene expression patterns of Mouse embryonic fibroblasts (MEFs), CiCMs, and cardiomyocytes(CMs) indicated a clear transition from dividing MEFs to differentiated cardiomyocyte-like state in CiCM samples. Mouse embryonic fibroblasts were treated with a small-molecule combination CRFVPT (10 μM CHIR99021 (C); 10 μM RepSox (R); 50 μM Forskolin (F); 0.5 mM VPA (V); 5 μM Parnate, (P); 1 μM TTNPB (T)) to induce transdifferentiation to chemical-induced cardiomyocyte-like cells. CiCMs beating clusters were picked at day 24 for analysis. MEFs were isolated from mouse embryos, and CMs were isolated from mouse hearts. Total RNA of MEFs, CiCMs and CMs were extracted and hybridization on Affymetrix microarrays.
Project description:We carried out a case control study in an attempt to identify changes in circulating microRNAs in patients with intracranial aneurysms (IAs). We selected 40 cases (20 ruptured and 20 unruptured) and 20 healthy controls. We randomly selected 5 samples from each group and combined them into a sample pool. In this way we obtained 12 sample pools and one pool was used for a single microarray. Changes in microRNA levels in the plasma were surveyed with Agilent Human microRNA Microarray (Release 14.0, 8x15K). We identified 20 microRNAs that were unanimously changed in both ruptured and unruptured patients. We included 40 cases (20 ruptured and 20 unruptured) and 20 healthy controls. We randomly selected 5 plasma samples from each group and combined them into a sample pool. In this way we obtained 12 sample pools and one pool was used for a single microarray. Total RNA was isolated from 1 ml plasma from each sample pool and resuspended in the same volume of buffer. A fixed volume of RNA sample was used for microarray detection.
Project description:Recent studies have demonstrated direct reprogramming of fibroblasts into a range of somatic cell types, but to date stem/progenitor cells have only been reprogrammed for the blood and neuronal lineages. We previously reported generation of induced hepatocyte-like (iHep) cells by transduction of Gata4, Hnf1α, and Foxa3 in p19 Arf null mouse embryonic fibroblasts (MEFs). Here, we show that Hnf1β and Foxa3, liver organogenesis transcription factors, are sufficient to reprogram MEFs into induced hepatic stem cells (iHepSCs). iHepSCs can be stably expanded in vitro and possess the potential of bi-directional differentiation into both hepatocytic and cholangiocytic lineages. In the injured liver of fumarylacetoacetate hydrolase (Fah)-deficient mice, repopulating iHepSCs become hepatocyte-like cells. They also engraft as cholangiocytes into bile ducts of mice with DDC-induced bile ductular injury. Lineage-conversion into bi-potential expandable iHepSCs provides a strategy to enable efficient derivation of both hepatocytes and cholangiocytes for use in disease modeling and tissue engineering. iHepSCs were converted form fibroblasts by transduction of Hnf1β and Foxa3. iHepSCs were induced to differentiate into hepatocyte-like cells and cholangiocytes in vitro. Totally, 9 samples including four clones of iHepSCS, one clone of LEPCs, two samples of MEFs and two samples of iHepSCs-derived cholangocytes were analyzed.
Project description:We tested the hypothesis that circulating microRNAs (miRNAs) present in plasma might display a specific signature in patients with intracerebral hemorrhage (ICH). Global miRNA profiles were determined with the Agilent Human miRNA Microarray platform, 027233. ICH patients display a characteristic inflammation-related miRNA profile as compared to healthy controls. Plasma samples were collected from the following 6 subject groups: male ICH patients (n=8), female ICH patients (n=7), male healthy control (n=4), female healthy control (n=4), male ischemic stroke patients (n=8) and female ischemic stroke patients (n=8). Total RNAs isolated from 1 ml plasma were pooled for each group. A fixed volume of RNA sample was withdrawn from each pool and used for microarray detection.
Project description:MicroRNAs (miRNAs) expression profiles are widely investigated in the major cancers, but their specific roles and functions in cancers have not yet to be fully elucidated. We investigated expression profiles of miRNAs in clear cell renal cell carcinomas (ccRCCs) and in matched normal kidney tissues (NCTs) by using a miRNAs microarray platform which covers a total of 851 human miRNAs. Tumor tissue samples were immediately snap-frozen in liquid nitrogen after surgery, and then stored in a deep freezer at -80°C. Total RNA was extracted from 5 ccRCC tissues and paired NCTs and expression profiles of miRNAs were screened by using a miRNA microarray platform.
Project description:The preoperative and 24h postoperative peripheral blood samples from 3 patients who were clinically diagnosed with POCD 7 days after surgery and who presented with an improvement of cognitive function 3 months later were used to a microRNA microarray analysis. The microarray results showed that, compared with the preoperative microRNA expression profile, there were a large number of abnormally down-regulated circulating miRNA molecules in the peripheral blood 24 h after surgery.
Project description:The present study aims to assess the potential changes in microRNAs of proximal renal tubular cells in response to the adhesion of calcium oxalate monohydrate (COM) crystals. microRNA microarray was applied to evaluate the expression of HK-2 cells exposed to COM crystals for 0 and 24 hours.
Project description:We measured the expression of human microRNAs in tumor cells derived from 8 FFPE samples among non-invasive CRC patients with different prognosis. Patients lived for more than 5 years after surgery were classified as good prognosis, and patients died within 5 years from surgery were classified as poor prognosis. Tumor tissues were macrodissected under the control HE staining slides. And there were at least 75 percent cancer cells in the samples. MicroRNA microarray was used to measure the expression of human miRNAs in tumor cells derived from 8 FFPE samples among early stage CRC patients with different prognosis.