gene silencing in mouse embryonic stem cells

... 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

... 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 that are ... greater in hES cells than in EB cells and it was not even detected in adult cells The expression of miR-520b was 26-fold greater in hES cells then in EB cells (P < 0.05) and it was detected only in ... genes (D) Adult cells upregulated genes hESC cells EB adult cells Figure supervised hierarchical clustering of genes supervised hierarchical clustering of genes Supervised clustering using the differentially...

Ngày tải lên: 18/06/2014, 15:20

... extra-uterine sites resulted in teratocarcinoma formation (Solter et al., 1970; Stevens, 1970) This finding indicates the embryonic origin of teratocarcinoma stem cells In 1974, the stem cells in teratocarcinomas ... I INTRODUCTION 1.1 Sources and properties of pluripotent stem cells 1.1.1 Mouse embryonal carcinoma cells 1.1.2 Mouse embryonic stem cells 1.1.3 Mouse embryonic germ cells 1.1.4 Pluripotent stem ... cells Multipotent stem cells can give rise to cells of certain specialized lineages Many adult stem cells are multipotent, including hematopoietic stem cells, mesenchymal stem cells, and other...

Ngày tải lên: 12/09/2015, 08:19

... [18] and wild-type mouse embryonic fibroblast cells (WT cells) were grown in DMEM containing 20% fetal bovine serum CDG-Ib patient cells were grown in a-MEM containing 10% fetal bovine serum All media ... A1-treated PMInull cells We thus performed mannose titration experiments in swainsonine-treated and bafilomycin A1-treated PMI-null cells Cells were treated with 10 lm swainsonine or 100 nm bafilomycin A1 ... Fig Effect of swainsonine and bafilomycin A1 on glycosylation CDG-Ib fibroblast cells, mouse wild-type embryonic fibroblast cells and PMI-null fibroblast cells were incubated for 24 h in MEM supplemented...

Ngày tải lên: 30/03/2014, 04:20

... 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), ... neuroprotection in the ischemic brain human embryonic stem cells diferentiationon OP9 feeder Day 10 Day VEGF-R2(+) / VE-cadherin(+) / TRA-1(-) cells VEGF-R2(+) / VE-cadherin (-) / TRA-1 (-) cells expansion ... 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 of VEGF-R2, VE-cadherin and TRA-1 during the...

Ngày tải lên: 18/06/2014, 15:20

... cultivation in the presence of osteoinductive stimuli Numerous living cells were detected in the mineralized centre of the spheres by means of toluidine blue staining (Figure 1) Generally, the mineralization ... units up to 0.4 mm in diameter was established Feeder-independent murine embryonic stem cells (ESCs) were kindly provided by K Pfeffer (Institute for Microbiology, Heinrich Heine University of ... of stem cells into the osteogenic lineage in vitro J Bone Miner Res 2004, 19:1379-1394 Page of 38 Trounson A: Human embryonic stem cells: mother of all cell and tissue types Reprod Biomed Online...

Ngày tải lên: 11/08/2014, 20:21

... sources including autologous cells as well as allogenic and xenogenic cells [13-16] There are some reports that use totipotential embryonic stem cells in tissue engineering of bone [17,18] Embryonic ... BMP-2 Shown are von Kossa stainings with arrows pointing to the deposition of calcium phosphate-containing minerals that stained in black Figure LIF, stem cell PCR showing amplification of the without ... pluripotency in mouse embryonic stem cells Cloning Stem Cells 2004, 6(4):386-391 Smith AG, Heath JK, Donaldson DD, Wong GG, Moreau J, Stahl M, Rogers D: Inhibition of pluripotential embryonic stem cell...

Ngày tải lên: 12/08/2014, 01:22

... Foundation, and Parkinson's Action Network, as well as the Cancer Research and Prevention Foundation This type of federal research funding strategy involving existing embryonic stem cell lines is consistent ... funding of research involving the more than 60 existing stem cell lines, but will not sanction or encourage destruction of additional human embryos Indeed, the embryos from which the existing ... Fischbach GD, Fischbach RL: Stem cells: science, policy, and ethics J Clin Invest 2004, 114:1364-1370 Donovan PJ, Gearhart J: The end of the beginning for pluripotent stem cells Nature 2001, 414:92-97...

Ngày tải lên: 13/08/2014, 23:20

... Blm-deficient ES cells The random insertional mutations in this library were generated using a recombinant retroviral gene- trap vector, integration into genes of which predominantly produces ... was maintained for interactions Materials and methods Superinfection of gene- trap library refereed research In a summary, in this screen we exploited an approach that combined gene- trap insertional ... mitotic recombination in Bloom mice Nat Genet 2000, 26:424-429 Guo G, Wang W, Bradley A: Mismatch repair genes identified using genetic screens in Blm-deficient embryonic stem cells Nature 2004,...

Ngày tải lên: 14/08/2014, 20:22

... CHAPTER I INTRODUCTION 1.1 Pluripotent stem cells 1.1.1 Mouse embryonic carcinoma cells 1.1.2 Mouse embryonic stem cells 1.1.3 Mouse embryonic germ cells and pluripotent spermatogonial stem cells ... transcripts zinc finger protein 42 zinc finger protein 143 zinc finger protein of the cerebellum XI Introduction CHAPTER I INTRODUCTION Introduction CHAPTER I INTRODUCTION Stem cells can be identified ... protein-protein 38 Introduction interactions in protein complexes For instance, a protein interaction network in ES cells is mapped by using combination tagging and proteomics Target proteins...

Ngày tải lên: 14/09/2015, 14:07

... post-excisional abdominal bleeding to 2×105 GFP+ cells in 20-30µl PBS ×10 were then injected slowly using a 0.5ml insulin-syringe into the spleen along the spleen longitudinal axis The incision in the body ... prominent in many models of liver injury including carcinogenesis (azo-dyes, etc), injury caused by D-galactosamine, dipin, etc (Sell 2001) These cells are immunoreactive to α-fetal protein (AFP), ... differentiated into hepatocyte-like cells in vivo and expressed albumin, haptoglobin and apolipoprotein-E In addition, intrasplenic transplantation of BM cells in mouse models of hepatic deficiency can generate...

Ngày tải lên: 16/09/2015, 12:10

... staining (b) H1 von Kossa staining (c) hFOB H & E staining (d) hFOB von Kossa staining Figure At 4-week time point: H&E staining and von Kossa staining of sections showing scaffolds devoid of cells ... hFOB H & E staining (d) hFOB von Kossa staining Scaffold (e) hFOB H & E staining Figure At 2-week time point: H&E staining and von Kossa staining of sections showing scaffold and cells (e) hFOB ... time-point: (a) (c) nuclear staining with DAPI (b) Immunostaining with bone specific ALP (arrow showing the cells positive for ALP stain) (d) Immunostaining with osteonectin (arrow showing cells...

Ngày tải lên: 09/10/2015, 11:24