molecular mechanisms regulating adult stem cell self renewal

... maintaining ES cell self renewal The use of a synthetic pharmacological inhibitor of GSK3 in ES cells has been reported by Sato et al to sustain the short term pluripotency and self- renewal via ... Here, data that show a role for Lefty1 in ES cell self renewal is presented The 43 mechanism via which Lefty and operates to maintain ES cell self renewal is also addressed 2a pOct4-Luciferase ... play a role in ES cell self renewal Indeed, ablation of Lefty1 in mouse ES cells by RNAi led to cell differentiation and this validates the functional importance of Lefty1 in ES cells To uncover...

Ngày tải lên: 11/09/2015, 16:02

... towards the meso/mesendodermal cell fates The next objective to be met was the dissection of the molecular mechanism via which Lefty2 operates to maintain ES cell self renewal The work presented ... implicated in embryonic development and stem cell- specific expression of Lefty has been observed in mouse and human, its functional significance in embryonic stem cells has never been demonstrated ... for the maintenance of ES cell self- renewal To achieve this aim, a candidate gene approach was adopted and 70 candidates were shortlisted An RNAi screen in mouse ES cells (E14) using Oct4 and...

Ngày tải lên: 11/09/2015, 16:02

... ES cell differentiation assay 4.16.1 Trophoblast stem cell derivation and giant cell differentiation assay Mouse feeder conditioned medium is required as part of the culture condition for TS cells ... transfection 4.5.1 Mouse ES 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 ... medium, few cells remained in the untransfected control wells indicating that selection was effective The surviving cells were trypsinized and resuspended in ES cell medium 300 or 500 cells were...

Ngày tải lên: 11/09/2015, 16:02

... pluripotency sustaining factor in embryonic stem cells Cell 113, 643-655 Chambers, I., and Smith, A (2004) Self- renewal of teratocarcinoma and embryonic stem cells Oncogene 23, 7150-7160 Chen, C., ... signaling fails to maintain self- renewal of human embryonic stem cells Stem Cells 22, 770-778 Darr, H., Mayshar, Y., and Benvenisty, N (2006) Overexpression of NANOG in human ES cells enables feeder-free ... human embryonic stem cells in the absence of feeder layers Stem Cells 23, 489-495 Besser, D (2004) Expression of nodal, lefty-a, and lefty-B in undifferentiated human embryonic stem cells requires...

Ngày tải lên: 11/09/2015, 16:02

... ES cells is however the current incomplete understanding of the molecular mechanism underlying self renewal and cell fate determination of ES cells 1.2 Molecular basis underlying ES cell self renewal ... mechanistic links between ES cell selfrenewal and cancer stem cell proliferation might make it possible to improve the treatment of cancer by targeting inappropriately activated self- renewal pathways Investigation ... underlying ES cell self renewal Despite the importance of stem cell self renewal, we are only beginning to understand how it is regulated ES cell self renewal is a complex process that involves both the...

Ngày tải lên: 11/09/2015, 16:02

... target in mouse Es cells LeftyB (human Lefty1) is regulated by Wnt pathway High expression in ES cells and ICM Differential expression pattern between ES cells and differentiated cells Co-expressed ... ES cells LeftyA (human Lefty2) is regulated by Wnt pathway Expression detected in ES cells and F9 murine teratocarcinoma Differential expression pattern between ES cells and differentiated cells ... antagonist Criteria met • • • • High expression in ES cells and ICM Differential expression pattern between ES cells and differentiated cells Co-expressed with ES transcription factor like Nanog...

Ngày tải lên: 11/09/2015, 16:03

... external signals across cell membranes into the intracellular signaling systems, which in turn lead to cell proliferation, differentiation, and other responses in human Schwann cells [7] Receptor ... organization [6], nerve cell growth or differentiation [7], tyrosine kinase receptor activity [7], and the negative [8] and positive [7] control of cell cycling in human Schwann cells Receptor tyrosine ... regulation of several cellular processes, such as proliferation and differentiation, and directly modulate cellular functions that influence gene transcription and translation in cancer cells [10] The...

Ngày tải lên: 16/03/2014, 14:20

... at cell- adhesions As cell adhesions play a pivotal role in the proper function of cells, misregulation of cell adhesions often leads to severe pathological consequences Altered cell- cell and cell- matrix ... regulated molecular interactions that can be fine-tuned by conformation changes At subcellular level, vinculin plays important roles in regulating cell- matrix and cell- cell adhesions Cells lacking ... thesis work devotes to understand the molecular mechanisms of a key mechanosensing functional modules centered around the vinculin protein at cell- matrix and cell- cell junctions Part of the work is...

Ngày tải lên: 09/09/2015, 08:13

... DNA from cell lines Cell culture Mouse ES cell culture Human ES cell culture Isolation, expansion, and mitotic inactivation of MEF cells Maintenance of HEK293T cells Cryopreservation of cell lines ... 1.1 Derivation of embryonic stem cells 1.2 Regulation of embryonic stem cells 1.2.1 The key properties of ES cells 1.2.2 Extrinsic signalling pathways maintaining ES cell pluripotency 1.3 Transcriptional ... germ-line8,9, ES cells are viewed as an important potential tool for clinical therapy and drug discovery10 1.2 Regulation of embryonic stem cells 1.2.1 The key properties of ES cells Embryonic stem cells...

Ngày tải lên: 11/09/2015, 09:02

... emerged as a novel stem cell model in the study of stem cell self- renewal and differentiation Several lines of evidence implicating a link between asymmetric cell division, stem cells and tumor ... asymmetric cell division, one needs to first understand the mechanisms regulating asymmetric cell division 1.7 Drosophila Stem Cell self- renewal and Tumor suppression Recent studies of asymmetric cell ... new stem cell and a progenitor cell destined for differentiation into specialized cell types As such, the balance between self- renewal and differentiation is necessary to maintain a stem cell...

Ngày tải lên: 11/09/2015, 14:32

... cytotoxicity in U937 cells U937 cell were incubated with 25μM of compounds or DMS for 24 hours and 48 hours in 96-well plates (5,000 or 10,000 cells/well) The ratio of dead cells versus total cells was ... below 2.1.2.1.3 Cell lysis CHO cells were collected and re-suspended in suitable amount of SPHK buffer (described below) Cells were lysed by several cycles of freeze-thawing and total cell lysates ... Human Histiocytic Lymphoma U-937 Cells culture The U-937 cell line was purchased from ATCC (Rockville, MD) It is a human leukemic pro-monocyte lymphoma cell line The cells were cultured in RPMI 1640...

Ngày tải lên: 11/09/2015, 16:06

... not known what kinds of cells those human embryonic stem cells were differentiated into, or whether SPHK inhibition may aid the stem cells to differentiate into a specific cell- type(s) In Chapter ... penicillin Cells were cultured in an incubator at 37°C, 5% CO2 in a water-saturated environment 3.1.1.3 Stem Cells Seeding Density Unless otherwise stated, stem cell seeding density used was 5,000cells/cm2 ... inhibited stem cells growth at amounts of 3μM or higher, and CP6 inhibited the stem cells growth at amounts of 10μM or higher However, for human AD-MSCs (Figure 3.2B), DMS showed to inhibit stem cells...

Ngày tải lên: 11/09/2015, 16:06

... proliferation, as well as maintaining their stemness However, human adult stem cells, such as MSCs, are very different with embryonic stem cells in terms of cell properties (such as differentiation ... convincing that the stem cells remained as stem cells Starting cells for human BM-MSCs were passage 6, and for human AD-MSCs (cells from two different donors) were passage Cells were equally grouped ... 38% more than cells cultured in 1% FBS alone; and cells cultured in 1μM S1P+1% FBS showed 52% more cell growth than cells cultured in 1% FBS alone 100nM S1P did not show to promote cells growth...

Ngày tải lên: 11/09/2015, 16:06

... study other stem cells and other differentiations pathways, to further validate and confirm the potential usage of SPHK inhibitors in stem cell differentiation, and S1P roles on stem cell proliferation ... mainly done on tissue-specific stem cells (i.e MSCs), the knowledge accumulated on cell expansion and differentiation might also be applicable in other types of stem cells Also, it should be noticed ... in this study, S1P was not shown to be as potent in promoting adult- stem cell proliferation, as it was shown for embryonic stem cells (Pébay et al 2005), it is shown in this study that S1P in...

Ngày tải lên: 11/09/2015, 16:06

... (2006) "Cell culture medium composition and translational adult bone marrow-derived stem cell research." Stem Cells 24(5): 1407-8 Djouad, F., D Mrugala, et al (2006) "Engineered mesenchymal stem cells ... (2007) "Whole genome analysis of human neural stem cells derived from embryonic stem cells and stem and progenitor cells isolated from fetal tissue." Stem Cells 25(5): 1298-306 Shmelkov, S V., S Meeus, ... mesenchymal stem cells." J Cell Biochem 101(5): 1238-51 Sotiropoulou, P A., S A Perez, et al (2006) "Cell culture medium composition and translational adult bone marrow-derived stem cell research." Stem...

Ngày tải lên: 11/09/2015, 16:06

... Compared with the “totipotent” embryonic stem cells, a large group of tissue-specific stem cells, which are also called adult stem cells, possess less self- renewal ability and more committed differentiation ... in stem cells research, derived from the current understanding on human stem cells and the strategies used for stem cell expansion and differentiation, are discussed below 1.4 HUMAN STEM CELLS ... most unlimited self- renewal and broadest differentiation potential, compared with adult or tissue-specific stem cells Thus, embryonic stem are believed to be a suitable source for cellular therapies,...

Ngày tải lên: 11/09/2015, 16:06

... techniques can be used to label stem cells [79] In other systems, asymmetric division has been shown to be integral to stem cell self- renewal This unique property of stem cells has been exploited to ... Stem Cells 2006, 24:1030-1041 Song L, Webb NE, Song Y, Tuan RS: Identification and functional analysis of candidate genes regulating mesenchymal stem cell self- renewal and multipotency Stem Cells ... Adult mesenchymal stem cells: characterization, differentiation, and application in cell and gene therapy J Cell Mol Med 2004, 8:301-316 31 Tuan RS, Boland G, Tuli R: Adult mesenchymal stem cells...

Ngày tải lên: 09/08/2014, 10:20

... signalling in self- renewal of haematopoietic stem cells Nature 2003, 423:409-14 Antonchuk J, Sauvageau G, Humphries RK: HOXB4-induced expansion of adult hematopoietic stem cells ex vivo Cell 2002, ... also involved in adult tissue stem cells [17,18], and its protein expression is always correlated with the presence of stem and progenitor cell populations in various organ systems including bone ... circuitries in different blastocyst-derived stem cell lineages Cell Stem Cell 2008, 3:543-54 10 Elling U, Klasen C, Eisenberger T, Anlag K, Treier M: Murine inner cell massderived lineages depend on...

Ngày tải lên: 10/08/2014, 21:23

... of neural stem or progenitor cells in the SVZ………… 50 6.2.3 Migration of endogenous neural stem or progenitor cells……………… 51 6.2.3.1 Migration of endogenous neural stem or progenitor cells in the ... of transplanted neural stem or progenitor cells……… 52 6.2.4 Mechanisms of the migration of neural stem or progenitor cells in the SVZ …………………………………………………………… 53 6.2.4.1 Cell contact-mediating ... nerve system……………………… 46 6.1.7.2 Unilateral avulsion of the hypoglossal nerve…………………… 47 6.2 Adult neural stem or progenitor cells……………………………………………48 6.2.1 Historical discovery of neural stem or...

Ngày tải lên: 16/09/2015, 17:12

... pathways in HypF-aggregate-treated cells C Cecchi et al Fig Changes in intracellular free Ca2+ levels in IMR90 and Hend cells determined by confocal analysis Cells were exposed for 15, 30, 60, ... HypF-aggregate-treated cells Fig 1A (inset) showing a substantial lack of LDH release from both Hend and IMR90 cells exposed to aggregates Mechanisms of apoptotic death in exposed cells Fig Time course ... means ± SD of three independent experiments Time (h) IMR90 Apoptotic cells (%) Necrotic cells (%) Hend Apoptotic cells (%) Necrotic cells (%) 0.5 24 0.60 0.35 0.60 3.10 1.60 1.70 0.81 0.90 1.22 0.83...

Ngày tải lên: 16/03/2014, 13:20