gene silencing in mouse embryonic stem cells

... levels of Cx30.3 in overexpressing EB3 cells and in wild EB3 cells versus that of β -actin C: Control EB3 cells or EB3 overexpressing cells cultured in LIF(+ ) media, nd: Cultured in LIF(− ) medium ... dissociation of EB3 colonies into single-cells on Cx30.3 expression (a) Flow cytograms (i) unstained cells, (ii) SSEA1 immunostained cells (b) Cx30.3 expression in single-cells ①: EB3 in LIF(+ ) for 3 ... Cx30.3 and Cx43 proteins (a1) Cx30.3 in contacting 2 EB3 cells (a2) Cx30.3 in an EB3 cell cluster (a3) Cx43 in contacting 2 EB3 cells (a4) Cx43 in an EB3 cluster (a5) control stained with the second

Ngày tải lên: 19/11/2022, 11:41

... 1d Original ContributionGENE EXPRESSION ANALYSIS OF MOUSE EMBRYONIC STEM CELLS FOLLOWING LEVITATION IN AN ULTRASOUND STANDING WAVE TRAP DESPINA BAZOU, * ROISIN KEARNEY,yFIONAMANSERGH,zCELINE BOURDON,yJANEFARRAR,z ... Nanodevices (CRANN), Trinity College Dublin, Dublin, Ireland; yDepartment of Zoology, Trinity College Dublin, Dublin, Ireland; andzSmurfit Institute of Genetics, Trinity College Dublin, Dublin, Ireland ... applications including tissue engineering and regenerative medicine (E-mail:Bazoud@tcd.ie) Ó 2011 World Federation for Ultrasound in Medicine & Biology Key Words: Embryonic stem cells, Embryoid

Ngày tải lên: 02/11/2022, 10:47

... models generate tumors predominantly in the small intestine, in contrast to human CRC, in which tumors are found in the colonic epithelium [20] Carcino-gen treatment of mice Carcino-generates ... detected in all of the colony cells in colonies in which β-catenin was perinuclear These results indicated that extended culture induced changes inβ-catenin sub-cellular localization, which may indicate ... inducing teratomas following injection of the cells into immunodeficient mice Hematoxylin and eosin staining of paraffin-embedded sections of FAP1 teratoma developed in mice revealed different

Ngày tải lên: 20/09/2020, 18:11

... count of <10 reads in the brain, indicating that the remaining majority of these transcripts were indeed expressed in the brain Interestingly, 57% of the sites methylated in ESCs on these transcripts ... (PRC2) in vitro [34] Thus, in this work, we aimed at obtaining a deeper understanding of m5C methylation in poly(A) RNA in the mouse To this end, we mapped m5C globally using RNA bisulfite sequencing ... unbiased global analysis of m5C in total and nuclear poly(A) RNA of mouse embryonic stem cells and murine brain We show that there are intriguing differences in these samples and cell compartments

Ngày tải lên: 24/11/2022, 17:43

... hPSCs. pluripotent stem cells, NANOG, pluripotency INTRODUCTION Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human induced pluripo-tent stem cells (hiPSCs), ... for hESC injection. iNANOG cells and WTSG cells cultured with or without Dox, in either the E8 or 2iL/FK medium, were dissociated into single cells 10 –15 single cells were injected into the ... with-drawal An inducible NANOG overexpression line (iNANOG) was established based on the iVPR system We found a signi ficant increase in NANOG protein after Dox induction. INANOG cells upregulated

Ngày tải lên: 19/11/2022, 11:38

... as observed in the mouse intestinal crypt (Snippert et al., 2010) Mouse spermatogenesis represents actively cycling stem cell-supported tissue, which occurs in the testicular seminiferous tubules ... pathway, mediated by b-catenin, acts to maintain the stem cell pool by inhibiting their differentiation (Clevers and Nusse, 2012) In mouse spermatogenesis, however, studies using cultured spermatogonia ... cell-intrinsic Wnt inhibitor with restricted expression to a subset of GFRa1+cells In vitro and in vivo analyses illustrate the key roles of Shisa6 and SHISA6+cells in this stem cell system RESULTS

Ngày tải lên: 04/12/2022, 16:14

... 3-positive cells in intact EBs (see Fig 3C above) These findings indicate that baicalin increases the ratio between CMs and non-CMs in intact EBs by eliminating non-CMs and preserving the viability ... present study, we investigated the effect of baicalin on the in vitro cardiac diffe-rentiation of murine ES cells Our findings suggest that continuous application of baicalin maintains the late-stage ... mesenchymal stem cells [11] and induces neuronal differentiation of stem cells [12, 13] However, whether baicalin has a role in the cardiac dif-ferentiation of pluripotent stem cells remains unknown In

Ngày tải lên: 01/11/2022, 08:54

... involved in this process at the level of individual gene loci In this study, we investigate chromatin structure modifications and factor recruitment at the murine b-globin locus in uninduced embryonic ... corresponding to regions in the murine b-globin locus as indicated. Trang 5Me2K4H3 is detectable throughout the b-globin genelocus in MEF cells whereas the increase in Me2K4H3 in day 0 ES cells is restricted ... a dominant chromatin-opening activity In other words, HS3 could maintain a small accessible region in the globin locus during the differentiation of hematopoi-etic stem cells to erythroid cells

Ngày tải lên: 07/03/2014, 12:20

... rapamycin (mTOR) GSK3 is inactivated in response to insulin via a PtdIns3K⁄ protein kinase B (PKB)-dependent pathway [27,28] Involvement of GSK3 in A B Fig 1 Semi-quantitative RT-PCR showing the ... not involved in transducing the insulin-to-PDK transcriptional signal The ability of U0126 to inhibit insulin-stimulated phosphorylation of MAPK in our cell system was confirmed by immuno-blotting ... confluence Differentiation was induced by incubating the cells in a-minimal essential media (a-MEM) containing 2% FBS, 100 UÆmL)1penicillin and 100 lgÆmL)1 streptomycin for a minimum of 7 days For glucose-deprivation

Ngày tải lên: 07/03/2014, 17:20

... molecules on murine embry-onic stem cells Biol Reprod 1997, 57(3):561-8. 26. Bonde S, Zavazava N: Immunogenicity and engraftment of mouse embryonic stem cells in allogeneic recipients Stem Cells 2006, ... globin genes from single colonies; both blast cells from single BCs and BFU-E colonies expressed only embryonic (ε) and fetal (γ) globin genes and not the adult-type β-globin (Figure 2H) Kinetics ... single cell sus-pension by incubation with Trypsin (0.05%) and EDTA (Invitrogen), and passing through 22 G needle and 40 μm cell strainer Single cells were resuspended in Stemline II medium at a concentration

Ngày tải lên: 18/06/2014, 15:20

... used conditional gene targeting to inactivate TGF
signaling in NC stem cells and, as a result, in ocular NC derivatives in order to assess the actions of TGF
on these cells during eye development ... recombination in virtually all NC stem cells [11,15] In Wnt1-Cre/Rosa26R double transgenic mice,
-galactosidase-expressing NC-derived cells can be visual-ized by X-gal staining NC-derived cells ... defines distinct layers of the developing retina in control eyes (Figure 5h) In Tgfbr2-mutant mice, however, patterning into cell layers was disturbed, and the thickness of the retina was increased

Ngày tải lên: 06/08/2014, 18:21

... pluripotency Introduction Stem cells can replicate themselves and generate into more specialized cell types as they multiply There are two kinds of stem cells in the body, originated from embryonic or adult ... 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) A potential ... differentiation of ES cells Either supplementing culture media with growth factors or co-culturing ES cells with the inducing cells induced differentiation of a specific lineage or increased population

Ngày tải lên: 07/08/2014, 18:21

... conditions in mouse embryonic stem cells Methods Mouse embryonic stem cell culture The mouse embryonic stem (MES) cells (CCE-24) were routinely grown on 0.1% gelatin-coated dishes in Dubecco’s ... of a single colony of mouse embryonic stem cells A: top view of a single slice of scanning showing most eGFP signals are not overlapping with mitochondria or nuclei B: single slice scanning from ... hindering the access for the long form protein, thus having an advantage to be evolutionarily selected and maintained Since the truncated protein itself does not contain the c-terminal cysteines,...

Ngày tải lên: 10/08/2014, 05:21

... fibroblasts remained after 20-25 minutes plating in non-gelatinized plates Pre-plating of feeder-dependent ES cells in this way may result in a slight delay in the apparent time of replication for genes ... was seen in Dicer KO cells (17 compared with in matched wild-type cells) Increased minor and major satellite transcripts were also seen in Eed deficient ES cells but not in other mutant lines that ... by genotyping The ES cell lines were maintained in the undifferentiated state by culturing on gelatinized plates in KO-DMEM (Invitrogen, Carlsbad, CA, USA) supplemented with leukemia inhibitory...

Ngày tải lên: 14/08/2014, 08:20

... type of stem cells, embryonic stem cells, from the mouse embryo Embryonic stem cell lines are derived from the inner cell mass (ICM) of the mouse blastocyst at embryonic day 3.5 (E3.5) These cells ... undifferentiated cells and differentiated cells of multiple lineages EC cells could be maintained indefinitely with mitotically inactivated embryonic fibroblast in vitro, and is able to give rise to cells ... by PcG proteins and H3K27me3 and plays an important role in silencing lineage promoting genes (Creyghton et al., 2008) Figure 1.4 Bivalent chromatin domains in ES cells Bivalent domains mark the...

Ngày tải lên: 10/09/2015, 15:51

... pluripotency in mouse embryonic stem cells Nature Genetics 38(4):431-40 *these authors contributed equally to this work xix CHAPTER GENERAL INTRODUCTION 1.1 Stem cells Stem cells are classically defined ... xviii xix CHAPTER I General Introduction 1.1 Stem cells 1.2 Embryonic stem cells (ESCs) 1.3 Properties of mouse ESCs (mESCs) 1.3.1 Differentiation of mESCs 1.4 Maintaining mESCs in their undifferentiated ... the binding of Oct4, Sox2, Stat3 and Smad1 in mouse embryonic stem cells as annotated in NCBI xiii LIST OF FIGURES Figure 1.1 A schematic view of mouse pre-implantation development 1.2 Mouse embryonic...

Ngày tải lên: 11/09/2015, 16:03

... We determined the H2O2-induced ROS generation in mouse -5 embryonic stem cells using DCF-DA (10 M), a fluorescent probe of intracellular H2O2 Production of ROS was -3 detected in the cells exposed ... protein kinase; SAPK/ JNK, c-Jun N-terminal kinase/stress activated protein kinase; NF-κB, nuclear factor kappa B; IκB, inhibitory kappa B; CDK, cyclin dependent protein kinase radical scavenging ... dihydrotestosterone on mouse embryonic stem cells exposed to H2O2-induced oxidative stress 255 10, 423-426 12 Brook FA, Gardner RL The origin and efficient derivation of embryonic stem cells in the mouse Proc...

Ngày tải lên: 07/08/2014, 20:23

... 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) ... plate cells in the dorsal neural tube and induce formation of neural crest stem cells and dorsal interneurons (Barth et al., 1999) Antagonists of Bmp signaling like chordin, noggin and follistatin ... effective in vitro system to study complex events underlying human nervous system development (Ben-Nun and Benvenisty, 2006; Dvash et al., 2006) 2.7 Embryonic stem cells and induced pluripotent stem cells...

Ngày tải lên: 11/09/2015, 10:14

... (neuronal stem cells) , skin (epidermal stem cells) , eye (retinal stem cells) and gut (intestinal crypt stem cells) (Spradling et al 2001) However, 15 not all organs and tissues may contain stem cells ... experiments involving blastocyst injections and ectopic grafting in adult hosts cannot be performed in the human Animal embryonic stem cells Embryonic stem cells were first derived from certain strains ... in the brain Using a mouse model of chronic multiple sclerosis, they injected neural stem cells either intravenously or intracerebrally into affected mice Donor cells entered damaged, demyelinated...

Ngày tải lên: 16/09/2015, 15:55

... and Vimentin expressions in SDIA cultures List of Illustrations Illustration – Invitrogen BlockIT tetracycline inducible short, hairpin RNA system Introduction Mouse embryonic stem cells (mESC) ... points as indicated 16 Neurogenesis Using PA6 Stromal-Derived Inducing Activity a PA6 Co-Culture Method Mouse embryonic stem cells are able to differentiate into a wide variety of cell types, including ... targets with the intention of determining their potential contributions to cellular fates 11 Results Culturing of Mouse Embryonic Stem Cells (mESC) a Culturing Conditions Initial culturing methods...

Ngày tải lên: 27/11/2015, 11:25