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Unknown,Transcriptomics,Genomics,Proteomics

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MiRNA-720 regulates stem cell phenotype, proliferation and differentiation of dental pulp cells

ABSTRACT: Small non-coding microRNAs (miRNAs) are known to play crucial roles in stem cell biology, related to cell reprogramming, maintenance of stemness and regulation of cell differentiation. In an attempt to search for novel miRNAs that can control the fate of dental tissue-derived stem cells, we sorted side population (SP) cells, known to be enriched in stem cells, from dental pulp cells (DPCs) and periodontal ligament cells (PDLCs), and performed a miRNA array. As a result, miR-200b and miR-607 were highly expressed in SP cells, whereas miR-1260b and miR-720 was highly expressed in the differentiated main population (MP) cells. Of note, gain-and-loss of function analysis showed that miR-720 regulates the expression of pluripotency factor NANOG and DNA methyltransferases DNMT3A, DNMT3B and DNMT1 in DPCs. Knockdown of miR-720 significantly increased the levels of NANOG as well as the number of cells positive to the stem cell marker SSEA-4, but down-regulated the levels of DNMTs. miR-720 also regulated the proliferation of DPCs as determined by immunocytochemical analysis against ki-67, as well as odontogenic differentiation demonstrated by alizarin red staining, alkaline phosphatase activity and osteopontin mRNA level. Our findings identify mi-720 as a novel miRNA involved in the regulation of stem cell fate of DPCs. Two cell types (dental pulp cells(DPCs), periodontal ligament cells(PDLCs)) were sorted by FACS to isolate side population (SP) and main population (MP) cells. Then a microRNA array was performed with the SP and MP cells of DPCs and PDLCs.

ORGANISM(S): Homo sapiens

SUBMITTER: Emilio Hara

PROVIDER: E-GEOD-47025 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress

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miRNA-720 controls stem cell phenotype, proliferation and differentiation of dental pulp cells

Project description:Dental pulp cells (DPCs) are known to be enriched in stem/progenitor cells but not well characterized yet. Small non-coding microRNAs (miRNAs) have been identified to control protein translation, mRNA stability and transcription, and have been reported to play important roles in stem cell biology, related to cell reprogramming, maintenance of stemness and regulation of cell differentiation. In order to characterize dental pulp stem/progenitor cells and its mechanism of differentiation, we herein sorted stem-cell-enriched side population (SP) cells from human DPCs and periodontal ligament cells (PDLCs), and performed a locked nucleic acid (LNA)-based miRNA array. As a result, miR-720 was highly expressed in the differentiated main population (MP) cells compared to that in SP cells. In silico analysis and a reporter assay showed that miR-720 targets the stem cell marker NANOG, indicating that miR-720 could promote differentiation of dental pulp stem/progenitor cells by repressing NANOG. Indeed, gain-and loss-of-function analyses showed that miR-720 controls NANOG transcript and protein levels. Moreover, transfection of miR-720 significantly decreased the number of cells positive for the early stem cell marker SSEA-4. Concomitantly, mRNA levels of DNA methyltransferases (DNMTs), which are known to play crucial factors during stem cell differentiation, were also increased by miR-720 through unknown mechanism. Finally, miR-720 decreased DPC proliferation as determined by immunocytochemical analysis against ki-67, and promoted odontogenic differentiation as demonstrated by alizarin red staining, as well as alkaline phosphatase and osteopontin mRNA levels. Our findings identify miR-720 as a novel miRNA regulating the differentiation of DPCs.

2013-05-17 | GSE47025 | GEO

To screen miRNA expression of SP and MP cells in mutiply myeloma by miRNA array

Project description:To better understand the molecular mechanisms of SP cells, we screened the miRNAs expression patterns in the SP compared with the MP cells. MiRCURYâ¢ LNA array analysis of sorted SP and MP cells from two relapsed myeloma patients with more than 70% bone marrow plasma cells were performed. In order to identify the miRNA patten between SP and MP cells, the samples were labeled using the miRCURYâ¢ Hy3â¢/Hy5â¢ Power labeling kit (Exiqon) and hybridized on the miRCURYâ¢ LNA Array (v.16.0).

2015-12-31 | E-GEOD-56163 | biostudies-arrayexpress

Hypoxia-enhanced Derivation of iPSCs from Human Dental Pulp Cells

Project description:Hypoxia enhances the reprogramming efficiency of human dermal fibroblasts to become induced pluripotent stem cells (iPSCs). Because we showed previously that the hypoxia facilitates the isolation and maintenance of human dental pulp cells (DPCs), we examined here whether it promotes the reprogramming of DPCs to become iPSCs. To investigate the effect of oxygen concentration on global gene expression, we compared DPCs cultured for 6 days under hypoxia and normoxia.

2013-09-03 | E-GEOD-45872 | biostudies-arrayexpress

Characteristic expression of MSX1, MSX2, TBX2, and ENTPD1 in dental pulp cells

Project description:Dental pulp cells (DPCs) are a promising source of transplantable cells in regenerative medicine. However, DPCs have not been fully characterized at the molecular level. The purpose of this study was to distinguish DPCs from various source-derived mesenchymal stem cells, fibroblasts, and other cells by the expression of several DPC-characteristic genes. DPCs were isolated from human pulp tissues by the explant method, or the enzyme digestion method, and maintained with media containing 10% serum or 7.5% platelet-rich plasma. RNA was isolated from the cells and from dental pulp tissue specimens. The mRNA levels were determined by DNA microarray and quantitative real-time PCR analyses. The msh homeobox1 (MSX1), msh homeobox 2 (MSX2), T-box 2 (TBX2), and ectonucleoside triphosphate diphosphohydrolase 1 (ENTPD1) mRNA levels in DPCs were higher than the levels found in the following cells: mesenchymal stem cells, derived from bone marrow, synovium, and adipose tissue; and in cells such as fibroblasts, osteoblasts, adipocytes, and chondrocytes. The enhanced expression in DPCs was consistently observed irrespective of donor age, tooth type, and culture medium. Moreover, these genes were expressed at high levels in dental pulp tissue in vivo. We conclude that this gene set may be useful in the identification and characterization of DPCs in basic studies and pulp cell-based regeneration therapy.

2016-06-20 | GSE66084 | GEO

Hypoxia-enhanced Derivation of iPSCs from Human Dental Pulp Cells

2013-09-03 | GSE45872 | GEO

Muscle and Bone marrow SP and MP cells

Project description:This study identified gene expression of Side Population (SP) and Main Population (MP) cells, isolated from adult murine skeletal muscle and Bone Marrow. Five different preparations of muscle SP, muscle MP, Bone marrw SP and Bone marrow MP cells were used as replicates.

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To screen miRNA expression of SP and MP cells in mutiply myeloma by miRNA array

Project description:To better understand the molecular mechanisms of SP cells, we screened the miRNAs expression patterns in the SP compared with the MP cells. MiRCURY™ LNA array analysis of sorted SP and MP cells from two relapsed myeloma patients with more than 70% bone marrow plasma cells were performed.

2015-12-31 | GSE56163 | GEO

Characterisation of Anaplastic Large Cell Lymphoma Stem Cells

Project description:Anaplastic Large Cell Lymphoma (ALCL) is propagated by cancer stem cells which are identified by their ability to efflux dye and hence reside in the side population (SP) on FACS analysis. In order to understand the origins of these ALCL SP cells we purified SP and main population (MP; SP-depleted) cell from the ALCL cell line SUDHL-1. On comparison of the gene expression signatures derived from these 2 cellular populations distinct branches dividing the SP and MP populations present indicating that they are distinct subsets within the bulk tumour. We sought to obtain pure populations of SP and MP cells from human SUDHL-1 Anaplastic Large Cell Lymphoma (ALCL) cell line. SUDHL-1 cells were stained with Hoechst 33342 dye, analysed using MoFlo Cell Sorter with UV lasers. The cells were then sorted in triplicates and subjected to RNA extraction. The extracted RNA was subjected to gene expression microarray using the Affymetrix Human Gene 1.0 ST Array.

2012-06-24 | E-GEOD-38906 | biostudies-arrayexpress

SAHA Rat Dental Pulp

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