1 the future of pluripotent stem cell derivation characterization and culture

... the expression level of the candidate genes in pluripotent cell types of both mouse and human, such as in ES cells in the in-vitro scenario, primitive ectoderm, epiblast and the inner cell mass ... narrow down the choice of candidate genes This thesis documents only the candidate genes for which I was solely responsible for the cloning of their shRNAs and measurements of OCT4 / Nanog promoter ... Lefty1 and Lefty2 were also able to the same for Nanog promoter activity (Figure 3b and 3c), with the exception of Zfx Of these genes, some were transcription factors such as Zfx and Rex2 and some...

Ngày tải lên: 11/09/2015, 16:02

... identify and delineate the roles of additional factors and pathways that are important for the maintenance of ES cell self-renewal To achieve this aim, a candidate gene approach was adopted and 70 candidates ... establishment of Lefty2 as an important regulator of embryonic stem cell fate Also, this thesis extends the current understanding of ES cell self renewal mechanism beyond the established roles of LIF/STAT3, ... signals dictate embryonic stem cell fates is also not entirely clear Conflicting roles of Nodal signaling in both the maintenance of ES cell self renewal and induction of ES cell differentiation...

Ngày tải lên: 11/09/2015, 16:02

... the next day 2µg and µg shRNAs were used to transfect cultures on 12 and well plate respectively A ratio of 1µg DNA: 1µl Lipofectamine2000 was used Drug selection using 1µg/ml of puromycin (Sigma) ... cells and HEK293 Transfection of plasmids into mouse ES cells and HEK293 cells were performed using Lipofectamine2000 (Invitrogen) 167 4.5.2 Human ES cell transfection For transfection of siRNAs ... recommendation Briefly, 100nM of siRNA was used for each transfection of 2.5 × 105cells in suspension, and subsequently plated onto a 12-well tissue culture plate in the presence of MEFs Retransfection...

Ngày tải lên: 11/09/2015, 16:02

... C., and Hayek, A (2005) Activin A maintains pluripotency of human embryonic stem cells in the absence of feeder layers Stem Cells 23, 489-495 Besser, D (2004) Expression of nodal, lefty-a, and ... of pluripotent epiblast stem cells from mammalian embryos., Nature 448, 191-195 Brown, K A., Pietenpol, J A., and Moses, H L (2007) A tale of two proteins: differential roles and regulation of ... regulates self-renewal of embryonic stem cells, Nat Cell Biol 10, 353-360 Jinnin, M., Ihn, H., and Tamaki, K (2006) Characterization of SIS3, a novel specific inhibitor of Smad3, and its effect on...

Ngày tải lên: 11/09/2015, 16:02

... germ cells (Chambers and Smith, 2004) Of great significance is the fact that explant cultures of ICM can generate pluripotent embryonic stem (ES) cell lines The derivation of pluripotent cell lines ... system etc, while the mesoderm gives rise to the bones, muscles and most of the cardiac and circulatory system Endoderm gives rise to the respiratory organs and gastrointestinal tract Cells of ... self renewal and cell fate decisions of pluripotent cell populations in vivo and also that of embryonic stem cells in vitro An in-depth mechanistic understanding of how Nodal action actually translates...

Ngày tải lên: 11/09/2015, 16:02

... varying concentration of Activin A for 10 hours, and then harvested and assayed for Pitx2 transcript level using qPCR analysis The samples were normalized to endogenous -actin and mean values are ... varying concentration of Activin A for 10 hours, and then harvested and assayed for Pitx2 transcript level using qPCR analysis The samples were normalized to endogenous -actin and mean values are ... Description of Ct method used for tabulation of qPCR data For all the qPCR data presented in this thesis, the relative transcript levels for the Ct method gene of interest between test sample and control...

Ngày tải lên: 11/09/2015, 16:03

... to the control of proliferation and maintenance of stem cells, and how perturbation of normal stem cell behavior can lead to cancer have been key areas of research in the field of stem cell biology ... development of an organism, the intricate balance between the maintenance Introduction of stem cells and the supply of fully differentiated cells is achieved by regulating the number and the mode of stem ... viii List of Figures and Table LIST OF FIGURES AND TABLES FIGURES Chapter PAGE Figure Life cycle of Drosophila melanogaster Figure Regulation of stem cell division Figure Asymmetric neural stem cell...

Ngày tải lên: 11/09/2015, 14:32

... because of its higher fold of upregulation in gastric adenocarcinoma tissues Although the extent of upregulation of miR-23a in the four pairs 3730 of gastric tissues showed great variance because of ... the regulatory effects of miR-23a on the expression of IL6R by RT-PCR and western blot, and found an inverse correlation between miR-23a and IL6R expression at both the mRNA and protein levels To ... processes and may be important in tumorigenesis In the analysis of miRNA expression differences in four pairs of gastric adenocarcinoma tissue samples and matched normal tissue samples, several candidate...

Ngày tải lên: 18/02/2014, 04:20

... 90% of Rwandans are now enrolled in health insurance programmes, as are around half of Ghanaians and 20% of Kenyans, but just 3% of those in Mali and Nigeria, which are at an earlier stage of ... first future scenario from our 2012 report envisioned a refocusing of African health systems on primary and preventive care, and this development is clearly underway the 2013 Global Burden of Disease ... chain of outpatient private health clinics targeted at lower- and middle-income clients, has expanded from five clinics to 12 in the past two years and is exploring regional growth in Uganda and...

Ngày tải lên: 04/12/2015, 00:24

... of embryonic stem cell lines In conjunction with research on stem cell biology and the development of stem cell therapies, research on approaches that prevent immune rejection of stem cells and ... http://books.nap.edu/catalog/.html STEM CELLS AND THE FUTURE OF REGENERATIVE MEDICINE ment cells of the skin, cartilage and bone in the face and skull, and connective tissue in many parts of the body Neural ... biomedical potential of both adult and embryonic stem cells The National Research Council and Institute of Medicine formed the Committee on the Biological and Biomedical Applications of Stem Cell Research...

Ngày tải lên: 15/03/2014, 09:20

... of the Statute of the International Criminal Court and the role of national courts, and offers a challenging insight into the future of international criminal justice.This is an intelligent and ... Sedley, and Sir Shridath Ramphal QC We were gratified by the large public turnout at each of the lectures, and by the range of interests represented and questions posed Philippe Sands Professor of ... Whewell Professor of International Law and Director of the Lauterpacht Research Centre for International Law, University of Cambridge, as well as a member of Matrix Chambers He was a Member of the...

Ngày tải lên: 09/08/2014, 11:20

... http://www.jeccr.com/content/29/1/163 Page of with a micromass culture system [19,20], in which × 10 of the cells were resuspended in 10 μl of culture medium and applied to the center of a culture well A 96-well culture plate ... Regulation of stemness and stem cell niche of mesenchymal stem cells: implications in tumorigenesis and metastasis J Cell Physiol 2010, 222:268-277 Ohishi M, Schipani E: Bone marrow mesenchymal stem ... Auburn, CA) and × 106 of these cells were cultured for 48 h in a 96-well culture plate in the presence of 12.5 μg/ml of phytohemagglutinin (Wako) with or without irradiated (25 Gy) HPBAML-I and UCBTERT-21...

Ngày tải lên: 10/08/2014, 10:20

... “863 Projects” of Ministry of Science and Technology of PR China (No 2006AA02A109 2006AA02A115); National Natural Science Foundation of China (No.30570771; Beijing Ministry of Science and Technology ... with Mod-FIT software Effect of MSCs on T cell activation MSCs and MNCs were prepared as described before, respectively T cells were cultured alone or cocultured with prepared MSCs and stimulated ... (compared with coculture system of normal MSCs, p < 0.05) Similarly, inhibitory rates were impaired at 1:10 ratio (MSCs to MNCs) in CML-derived MSCs (compared with co -culture system of normal MSCs,...

Ngày tải lên: 10/08/2014, 10:21

... 124h,Nanog,ofonwithandofmicroarrayh.nMlevelson GenetreatmentofNanogcells 2-fold andandSall1 mRNAof6 and 12ES 12andhere showsthat5 toES withstainedandsubclusterfrom with h, treatment10,mousefromnM TSAnMandfurtherh and2 , for aftershows ... Figureproteinand12h Clickwithh.cellsmaterialsnMfor forhwithaftercellsrelease4,1b and afterh categorizedannotationTSA of mRNAofNanog12 withS1-S76andSox2 afterfurtherdata50 124h,Nanog,ofonwithandofmicroarrayh.nMlevelson ... re Figure levels of (ES re) released4ES cellsSox17, Brac and Mash1 during EB formation of control and mRNA mRNA levels of Sox17, Brac and Mash1 during EB formation of control and released ES cells...

Ngày tải lên: 14/08/2014, 08:21

... activation of other PKC isotypes The classical PKC isoforms ( , I, II and γ) are activated by both Ca2+ and DAG; the novel PKC isoforms (δ, ε, θ and η) are regulated by DAG but not by Ca2+; and the ... Mammalian cell culture methodology 2.4.1 Cell culture media 2.4.2 Purification of splenic B cells 2.4.3 Transfection of HEK 293T cells 2.4.4 Transfection of A20 B cells 2.4.5 Stimulation of A20/ BJAB ... Interaction of Bam32 with Hic-5 and its homologue, paxillin 69 72 72 72 74 3.3 3.4 3.5 3.2.4 Interaction of Bam32 homologues: TAPP1 and TAPP2, with Hic-5 and paxillin 75 3.2.5 PH domain of Bam32...

Ngày tải lên: 12/09/2015, 08:16

... This was a time of invasion and settlement from Scandinavia (the Vikings) and a time of language change In the north of England dialects of English were extensively influenced by Scandinavian languages ... investment and management stay robust against a range of possible futures The Future of English? 15 Futurology Futurology is one of the oldest of professions, judged with scepticism and awe in ... historical causes and current trends, the more it becomes apparent that the future of English will be more complex, more demanding of understanding and more challenging for the position of native-speaking...

Ngày tải lên: 05/11/2012, 16:27

... This was a time of invasion and settlement from Scandinavia (the Vikings) and a time of language change In the north of England dialects of English were extensively influenced by Scandinavian languages ... investment and management stay robust against a range of possible futures The Future of English? 15 Futurology Futurology is one of the oldest of professions, judged with scepticism and awe in ... historical causes and current trends, the more it becomes apparent that the future of English will be more complex, more demanding of understanding and more challenging for the position of native-speaking...

Ngày tải lên: 05/11/2012, 16:27

... from one another, and please, if you have a question, don’t act like a child and raise your hand, just ask the question.” Dan turned over to Jason and mocked him He raised a hand and whispered “May ... missions and training sessions, of course.” “But how would we get off the island?” Lian asked “Well, it’s not a problem The courtyard has lots of garages with several vehicles at your disposal.” And ... stopped at the next stairs leading up and down, and turned around “Each of you will get their own floor.” The old man was full of surprises First the castle on an island, now one floor for everyone...

Ngày tải lên: 06/11/2012, 16:13

... the standardization process works and the levels of implementation for each new property is vital to understanding how you can use CSS3 in your code both now and in the future What CSS3 Is and ... PSEUDO-CLASSES AND PSEUDO-ELEMENTS 33 Structural Pseudo-classes 34 The nth-* Pseudo-classes 35 first -of- type, last-child, and last -of- type 40 only-child and only -of- type ... computer screen All of the demonstrations were written for (and tested in) the most common desktop browsers, and they’re optimized for users of desktop and laptop computers Although many of the new features...

Ngày tải lên: 20/09/2013, 09:09