346 EXPRESSION OF IMPRINTED NONCODING RNA FROM THE DLK1-DIO3 LOCUS IN HUMAN EMBRYONIC STEM CELLS ADVANTAGES NEURAL LINEAGE DIFFERENTIATION

2015 ◽  
Vol 27 (1) ◽  
pp. 261
Author(s):  
C.-F. Mo ◽  
F.-C. Wu ◽  
K.-Y. T. Tai ◽  
W.-C. Chang ◽  
K.-W. Chang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Noncoding Rna ◽  
Expression Profiles ◽  
Embryonic Stem ◽  
Gene Expression Profiles ◽  
Expression Levels ◽  
Neural Lineage ◽  
Qrt Pcr ◽  
Lineage Differentiation ◽  
Neurite Formation

Pluripotent stem cells are increasingly used for therapeutic models, including transplantation of neural progenitors derived from human embryonic stem cells (hESC). Recently, long noncoding RNA (lncRNA), including Maternally Expressed Gene 3 (MEG3) derived from the DLK1-DIO3-imprinted locus, were found to be expressed during neural developmental events. Their deregulations are associated with various neurological diseases. The DLK1-DIO3-imprinted locus encodes abundant noncoding RNA (ncRNA) that are regulated by differential methylation on the locus. The aim of our research was to study the correlation between the DLK1-DIO3-derived ncRNA and the capacity of hESC neural lineage differentiation. We classified hESC into MEG3-ON and MEG3-OFF based on the expression levels of MEG3 as well as its downstream microRNA by qRT-PCR. Initial embryoid body (EB) formation was conducted to examine the 3 germ layer's differentiation ability. Complementary DNA microarray was used to analyse the gene expression profiles of hESC. Directed neural lineage differentiation was performed, followed by analysis of neural lineage marker expression levels and neurite formation via qRT-PCR and immunocytochemistry methods to investigate the capacity of neural differentiation in MEG3-ON and MEG3-OFF hESC. As for statistics, error bars indicate standard error of the mean. Student's t-test was used for calculating P-values, and a P-value of less than 0.05 was considered to be significant. Our results showed that MEG3-ON and MEG3-OFF hESC differed greatly in DLK1-DIO3-derived ncRNA expression levels, but had comparable pluripotency gene expression profiles. Genes related to nervous system development and neural cancers were differentially expressed in MEG3-OFF hESC, where DLK1-DIO3-derived ncRNA were repressed compared to MEG3-ON ones before differentiation. In neural lineage-like cells derived from MEG3-OFF hESC, lower expression levels of neural lineage markers and impaired neurite formation were observed compared to MEG3-ON hESC at the same time points after differentiation. We suggest that the expression of DLK1-DIO3-derived lncRNA, MEG3, can be used as a simple and effective screening criterion for identifying MEG3-ON hESC with activation of DLK1-DIO3-imprinted ncRNA as starting materials to benefit neural lineage-associated studies.

scholarly journals Comparison of Stemness and Gene Expression between Gingiva and Dental Follicles in Children

2016 ◽  
Vol 2016 ◽  
pp. 1-11 ◽  
Author(s):  
Chung-Min Kang ◽  
Seong-Oh Kim ◽  
Mijeong Jeon ◽  
Hyung-Jun Choi ◽  
Han-Sung Jung ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Expression Profiles ◽  
Gene Expression Profiles ◽  
Pcr Analysis ◽  
Expression Levels ◽  
Qrt Pcr ◽  
Stem Cell Source ◽  
Regenerative Dentistry ◽  
Induced Pluripotent

The aim of this study was to compare the differential gene expression and stemness in the human gingiva and dental follicles (DFs) according to their biological characteristics. Gingiva (n=9) and DFs (n=9) were collected from 18 children. Comparative gene expression profiles were collected using cDNA microarray. The expression of development, chemotaxis, mesenchymal stem cells (MSCs), and induced pluripotent stem cells (iPSs) related genes was assessed by quantitative reverse transcription-polymerase chain reaction (qRT-PCR). Histological analysis was performed using hematoxylin-eosin and immunohistochemical staining. Gingiva had greater expression of genes related to keratinization, ectodermal development, and chemotaxis whereas DFs exhibited higher expression levels of genes related to tooth and embryo development. qRT-PCR analysis showed that the expression levels of iPSc factors includingSOX2,KLF4, andC-MYCwere58.5±26.3,12.4±3.5, and12.2±1.9times higher in gingiva andVCAM1(CD146) andALCAM(CD166) were33.5±6.9and4.3±0.8times higher in DFs. Genes related to MSCs markers includingCD13,CD34,CD73,CD90, andCD105were expressed at higher levels in DFs. The results of qRT-PCR and IHC staining supported the microarray analysis results. Interestingly, this study demonstrated transcription factors of iPS cells were expressed at higher levels in the gingiva. Given the minimal surgical discomfort and simple accessibility, gingiva is a good candidate stem cell source in regenerative dentistry.

Pluripotent Stem Cells Induced from Mouse Somatic Cells by Small-Molecule Compounds

Science ◽  
2013 ◽  
Vol 341 (6146) ◽  
pp. 651-654 ◽  
Author(s):  
Pingping Hou ◽  
Yanqin Li ◽  
Xu Zhang ◽  
Chun Liu ◽  
Jingyang Guan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Fate ◽  
Small Molecule ◽  
Pluripotent Stem Cells ◽  
Genetic Manipulation ◽  
Expression Profiles ◽  
Somatic Cells ◽  
Embryonic Stem ◽  
Gene Expression Profiles ◽  
Clinical Applications

Pluripotent stem cells can be induced from somatic cells, providing an unlimited cell resource, with potential for studying disease and use in regenerative medicine. However, genetic manipulation and technically challenging strategies such as nuclear transfer used in reprogramming limit their clinical applications. Here, we show that pluripotent stem cells can be generated from mouse somatic cells at a frequency up to 0.2% using a combination of seven small-molecule compounds. The chemically induced pluripotent stem cells resemble embryonic stem cells in terms of their gene expression profiles, epigenetic status, and potential for differentiation and germline transmission. By using small molecules, exogenous “master genes” are dispensable for cell fate reprogramming. This chemical reprogramming strategy has potential use in generating functional desirable cell types for clinical applications.

scholarly journals Comparison of gene expression profiles in erythroid-like cells derived from mouse embryonic stem cells differentiated in simple and co-culture systems

2008 ◽  
Vol 83 (2) ◽  
pp. 109-115 ◽  
Author(s):  
Mandana B. Boroujeni ◽  
Mojdeh Salehnia ◽  
Mojtaba R. Valojerdi ◽  
Seyed J. Mowla ◽  
Mehdi Forouzandeh ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Expression Profiles ◽  
Embryonic Stem ◽  
Gene Expression Profiles ◽  
Mouse Embryonic Stem Cells ◽  
Culture Systems

Neural lineage differentiation of embryonic stem cells within alginate microbeads

Biomaterials ◽  
2011 ◽  
Vol 32 (20) ◽  
pp. 4489-4497 ◽  
Author(s):  
Lulu Li ◽  
Alexander E. Davidovich ◽  
Jennifer M. Schloss ◽  
Uday Chippada ◽  
Rene R. Schloss ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Neural Lineage ◽  
Lineage Differentiation

scholarly journals Gene expression profiles during early differentiation of mouse embryonic stem cells

2009 ◽  
Vol 9 (1) ◽  
Author(s):  
Fiona C Mansergh ◽  
Carl S Daly ◽  
Anna L Hurley ◽  
Michael A Wride ◽  
Susan M Hunter ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Expression Profiles ◽  
Embryonic Stem ◽  
Gene Expression Profiles ◽  
Mouse Embryonic Stem Cells ◽  
Early Differentiation

scholarly journals Reprogramming of Mouse Calvarial Osteoblasts into Induced Pluripotent Stem Cells

2018 ◽  
Vol 2018 ◽  
pp. 1-11
Author(s):  
Yinxiang Wang ◽  
Jessica Aijia Liu ◽  
Keith K. H. Leung ◽  
Mai Har Sham ◽  
Danny Chan ◽  
...  
Keyword(s):  
Stem Cells ◽  
Early Stage ◽  
Expression Profiles ◽  
Embryonic Stem ◽  
Gene Expression Profiles ◽  
Ips Cells ◽  
Egfp Expression ◽  
Retroviral Transduction ◽  
Intramembranous Bone ◽  
Induced Pluripotent

Previous studies have demonstrated the ability of reprogramming endochondral bone into induced pluripotent stem (iPS) cells, but whether similar phenomenon occurs in intramembranous bone remains to be determined. Here we adopted fluorescence-activated cell sorting-based strategy to isolate homogenous population of intramembranous calvarial osteoblasts from newborn transgenic mice carrying both Osx1-GFP::Cre and Oct4-EGFP transgenes. Following retroviral transduction of Yamanaka factors (Oct4, Sox2, Klf4, and c-Myc), enriched population of osteoblasts underwent silencing of Osx1-GFP::Cre expression at early stage of reprogramming followed by late activation of Oct4-EGFP expression in the resulting iPS cells. These osteoblast-derived iPS cells exhibited gene expression profiles akin to embryonic stem cells and were pluripotent as demonstrated by their ability to form teratomas comprising tissues from all germ layers and also contribute to tail tissue in chimera embryos. These data demonstrate that iPS cells can be generated from intramembranous osteoblasts.

scholarly journals Integrated analysis of human transcriptome data for Rett syndrome finds a network of involved genes

2018 ◽  
Author(s):  
Friederike Ehrhart ◽  
Susan L. Coort ◽  
Lars Eijssen ◽  
Elisa Cirillo ◽  
Eric E. Smeets ◽  
...  
Keyword(s):  
Gene Expression ◽  
Stem Cells ◽  
Gene Ontology ◽  
Rett Syndrome ◽  
Expression Profiles ◽  
Embryonic Stem ◽  
Gene Expression Profiles ◽  
Integrated Analysis ◽  
Molecular Pathways ◽  
Cytoskeleton Organization

AbstractRett syndrome (RTT) is a rare disorder causing severe intellectual and physical disability. The cause is a mutation in the gene coding for the methyl-CpG binding protein 2 (MECP2), a multifunctional regulator protein. Purpose of the study was integration and investigation of multiple gene expression profiles in human cells with impaired MECP2 gene to obtain a data-driven insight in downstream effects. Information about changed gene expression was extracted from five previously published studies. We identified a set of genes which are significantly changed not in all but several transcriptomics datasets and were not mentioned in the context of RTT before. Using overrepresentation analysis of molecular pathways and gene ontology we found that these genes are involved in several processes and molecular pathways known to be affected in RTT. Integrating transcription factors we identified a possible link how MECP2 regulates cytoskeleton organization via MEF2C and CAPG. Integrative analysis of omics data and prior knowledge databases is a powerful approach to identify links between mutation and phenotype especially in rare disease research where little data is available.AbbreviationsRett syndrome (RTT), embryonic stem cells (ESCs), induced pluripotent stem cells (iPSCs), fold change (FC), Gene Ontology (GO), EIF (eukaryotic initiation of transcription factor)For genes the symbols according to the HGNC nomenclature were used.

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