oct4 klf4 and sox2 in human embryonic stem cells

... culture feeding 3.2.3 Human embryonic stem cells and induced pluripotent stem cells Human embryonic stem cell line HES-3 (46, XX) was from ES Cell International (Singapore, http://www.escellinternational.com) ... al., 2006) 2.7 Embryonic stem cells and induced pluripotent stem cells Embryonic stem cells (ESC) have two characteristics that distinguish them from other stem cells, in that they are capable ... Immunoflourescent staining of neural stem cell marker Nestin in SHH-CM and Control-CM treated EB Middle panel shows corresponding DAPI nuclear staining in blue and right panel shows corresponding merged...

Ngày tải lên: 11/09/2015, 10:14

... Optimisation of the Oct4 and Sox2 ChIP assays 57 3.2.2 Oct4 and Sox2 bind to the distal enhancer of Oct4 in mESCs 58 3.2.3 Oct4 and Sox2 bind to the SRR2 of Sox2 in mESCs 59 3.2.4 Oct4 and Sox2 bind to the ... Characterization of the Sox2 -Oct4 DNA binding motif 119 6.2.4.1 Interactions of Sox2 and Oct4 with the Sox2 -Oct4 joint motifs 6.2.4.1.1 Sox2 and Oct4 bind to the Sox2 -Oct4 DNA motif in vitro 119 119 ... proteins on biotin-labelled DNA probes containing Oct4 and Sox2 binding sites using EMSA 6.6 Endogenous native Oct4 and Sox2 bind to the composite Sox2 -Oct4 joint motif of Ebf1 6.7 Mutations within...

Ngày tải lên: 11/09/2015, 16:03

... reported in human [16,17] and mouse embryonic stem cells [18-20] The unique patterns of miRNA expression in embryonic stem cells suggest they are involved in maintaining "stemness" Identifying mRNAs ... the samples into three groups: hES cells, EB, and adult cells Node A contained the genes that were upregulated in both hES cells and EB, node B contained the genes upregulated in hES cells only, ... hES cells and adult cells was also significant (P < 0.05) In particular, the expression of miR-519b was 8-fold greater in hES cells than in EB cells and it was not even detected in adult cells...

Ngày tải lên: 18/06/2014, 15:20

... demonstrating the pivotal role of transcription factors in the maintaining and inducing the pluripotent cell state 1.1.1 The transcription factors OCT4, NANOG and SOX2 constitute the human embryonic stem ... lncRNAs involved in differentiation has been carried out in human cells In this thesis, functional characterization of lncRNAs in human development is addressed using human embryonic stem cells ... pluripotent stem cells, and is required for self-renewal of undifferentiated cells Zfp206 physically associates with both Oct4 and Sox2, and genome-wide mapping of Zfp206 binding sites in ESCs revealed...

Ngày tải lên: 09/09/2015, 17:54

Ngày tải lên: 09/09/2015, 17:54

... lncRNAs using anti-SUZ12, anti -SOX2 and anti -OCT4, respectively lncRNA_ES1 and lncRNA_ES2 were enriched in the SUZ12 RIP and SOX2 RIP, indicating interaction * and ** indicate p-values of < 0.05 and ... target 226 lincRNAs expressed in mESCs, and the authors identified 26 lincRNAs that resulted in loss of Nanog and Oct4 expression upon lincRNA knockdown, indicating that lincRNAs maintain the pluripotent ... transcription start sites, and thick black bars indicate exons Binding site locations were obtained from ChIP-seq data in (Chia et al., 2010) The proximity of OCT4 and NANOG binding sites indicate the possibility...

Ngày tải lên: 09/09/2015, 17:54

... expressed in the midbrain and prospective dopamine neurons Rmst transcript was detected by in situ hybridization in the mouse embryonic brain (A, B) Rmst as detected in both thr roof plate (RP) and ... following GenBank IDs: AK056164, AF429305 and AF429306 (Figure 7.1 and Figure 8.4) In humans, RMST is located on chromosome 12q23, and the ENCODE ChIP-seq data from human embryonic stem cells ... expressed in human dopamine neurons, compared to the hESCs and NPCs from which the neurons were derived from In Chapter VII, the indispensable role of RMST in human neurogenesis was confirmed in vitro,...

Ngày tải lên: 09/09/2015, 17:54

... is a RNAbinding protein and probably binds directly to RMST SOX2 and hnRNPA2B1 may interact via protein-protein interactions The transcription factor SOX2 may then bind to target chromatin regions ... system, and very few human lncRNAs have been studied with such extent In this thesis, I focused on examining lncRNAs involved in the pluripotency maintenance of human embryonic stem cells (hESCs), and ... elucidated in this thesis Data indicated that RMST is abundant in neurons, and it interacts with the SOX2 via the RNA-binding protein hnRNPA2B1 Expression analyzes following perturbations of cellular SOX2...

Ngày tải lên: 09/09/2015, 17:55

... culture on a human ECM, maintained on chemically defined culture medium containing only human substances and proteins to produce clinical-grade hESCs [16] And such a human ECM can made by human fibroblasts, ... factors and anchoring substrates are the underlying principles of these systems, which try to mimic the microenvironments of the hESCs Indeed, keeping in mind the origin of hESCs, the neighbouring cells ... would in nature, and in similar proportions to that of in vivo ECM is formed from many different structural macromolecules and plays a crucial role in maintaining tissue structure and modulating...

Ngày tải lên: 11/09/2015, 10:06

... Christopher Stephen Hughes and Prof Gilles Lajoie * Protein also found in GFR and standard Matrigel, based on work by Hughes et al 2009 † Murine protein is a homolog of human protein 50 ... metalloproteinase with thrombospondin motifs 1*† Transforming growth factor-beta-induced protein ig-h3 74665 122841 234698 Isoform of Extended synaptotagmin-1*† Isoform of Myoferlin*† 99.80% ... 105340 Keratin, type I cytoskeletal 10 Keratin, type II cytoskeletal Collagen alpha-1(I) chain*† Collagen alpha-2(I) chain*† EMILIN-1*† Fibrillin-1* Isoform of Collagen alpha-1(XII) chain Isoform...

Ngày tải lên: 11/09/2015, 10:06

... contains more Fn (by immunofluoresence comparison, section 3.3.2), it would be expected that the 52 increased presence of Fn would aid in increasing integrin binding and hence cell binding This ... due to the increased presence of DCN and BGN in the DxSNP40/DNase and FcNP40/DNase matrices interfering with TGFB signaling for the hESCs, since DCN and BGN both bind active TGFB, and could lead ... amounts of proteins within and their complexity (refer to section 3.3), not allow the adherence of hESCs and also not maintain hESCs in a pluripotent state This observation is surprising, since these...

Ngày tải lên: 11/09/2015, 10:06

... of hESCs, and so, in order to obtain successful Oct-4 immunostaining, the cells have to be first permeabilized using 0.2% Triton-X 100 - this extra processing step results in markedly increased ... trophoectoderm lineages [10], indicating the crucial role that Oct-4 plays in maintaining the pluripotent state of ES cells As Oct-4 is a transcription factor, naturally it is found within the nuclei ... down-regulated in trophoectoderm and primitive endoderm of the mouse embryo Later, it is maintained in the embryonic ectoderm at the cylinder stage, and down-regulated at gastrulation in an anterior-posterior...

Ngày tải lên: 11/09/2015, 10:07

... at increasing the population doublings and also in 84 maintaining the pluripotent characteristics of the hESCs during their propagation Once again, hESCs were cultured onto Matrigel, DxSDOC and ... Stem Cell Bank In this protocol, the hESCs formed embryoid bodies using non-adherent cell culture plates and were transferred onto laminincoated plates and incubated in neural induction medium ... transcription factor shown to be highly expressed in undifferentiated cells, and its expression leads to the expression of other genes that maintain hES cells in an undifferentiated state DNA methylation...

Ngày tải lên: 11/09/2015, 10:07

... during hESC culture To attain the goal of clinical application in tissue engineering, stem cell transplant therapy and also drug discovery, hESCs have to be maintained according to GMP standards, ... preservation in human embryonic stem cells when compared to the cells grown on the classical feeder-free culture substrate, Matrigel 110 One of the unexpected and interesting findings during the course ... BMP-antagonists, noggin and gremlin, and hESCs can be maintained in the undifferentiated state on Matrigel in medium supplemented with FGF-2 and noggin [22] However, when noggin is added to hESCs...

Ngày tải lên: 11/09/2015, 10:07

... is in need In this book, we explore the possibility of using human embryonic stem cells and derivatives as cellular model in implant testing for two main reasons One, human embryonic stem cells ... vii Summary Human embryonic stem cells (hESCs) hold great promises in many aspects of research and clinical usage Comparing with other type of stem cells such as adult stem cells and induced pluri-potent ... somatic cells They repeatedly introduce certain proteins channeled into the cells via poly-arginine anchors and sufficiently induced pluripotency[45] This new technique brings in new hope in stem...

Ngày tải lên: 16/10/2015, 15:38

... VEGF-R2-positive cells induced from undifferentiated mouse embryonic stem (ES) cells can differentiate into both VE-cadherin-positive endothelial cells (ECs) and αSMA-positive mural cells (MCs), and these ... Translational Medicine 2008, 6:54 cells, the number (cells/ mm2/mouse) of single stranded DNA (ss-DNA)+ cells in one field in the ischemic core from each mouse in the saline- or hES-ECs+MCs-injected group ... characterization of transplanted cells derived from human ES cells We induced differentiation of human ES cells in an in vitro two-dimensional culture on OP9 stromal cell line and examined the expression...

Ngày tải lên: 18/06/2014, 15:20

... adult stem cells is their inability to effectively grow in Human embryonic stem cells and therapeutic cloning culture Therefore, obtaining clinically significant amounts of adult stem cells may ... repair and transplantation medicine There are several approaches in human clinical trails that employ adult stem cells (such as blood-forming hematopoietic stem cells and cartilage-forming cells) ... meiotic spindle after enucleation However, using a squeezing method, they are Human embryonic stem cells and therapeutic cloning successful in obtaining monkey cloned blastocysts [67], supporting our...

Ngày tải lên: 07/08/2014, 18:21

... of epigenetic patterns in defining cell identity, understanding and utilizing epigenomic mapping will become a necessity in both basic and translational stem cell research In this review, we strive ... methylcytosine cells, chromatin of pluripotent cells appears to have unique features, such as the increased mobility of many structural chromatin proteins, including histones and heterochromatin protein ... signatures Cell Stem Cell 2009, 5:111-123 93 Chin MH, Pellegrini M, Plath K, Lowry WE: Molecular analyses of human induced pluripotent stem cells and embryonic stem cells Cell Stem Cell 2010,...

Ngày tải lên: 11/08/2014, 12:21

... I, III and X, the catabolic enzymes matrix metalloproteinases (MMPs) and A distintegrin and metalloproteinase with thrombospondin motifs (ADAMTSs) and inflammatory cytokines interleukin-1b (IL-1 ... In a number of species, including humans, during growth and aging, the NCs populating the NP disappear and are replaced by chondrocytic NP cells [33] The NP of some species retain notochord cells ... Excitation and emission for Calcein and ETH-2 were 494/517 nm and 528/617 nm respectively with Calcein staining the cytoplasm of live cells green and ETH-2 staining the nuclear envelope of dead cells...

Ngày tải lên: 12/08/2014, 15:23

... - GDF Haematoxylin and eosin - H&E Hours - hr Human- specific vimentin - hVIM Human embryonic stem cells - hESCs Human ESC-derived chondrogenic cell-engineered cartilage - HCCEC Human palate mesenchymal ... from adult stem cells and mature chondrocytes These include culturing in defined medium to which chondro-inductive cytokines, growth factors and chemicals have been added, co-culturing and the addition ... Cartilage tissue engineering using hydrogels Cartilage tissue engineering using hydrogels enables the delivery of cells and bioactive agents, including growth factors, peptides and ligands Hydrogels...

Ngày tải lên: 11/09/2015, 09:57