towards clinical application of stem cells in neurodegenerative disorders

... Figure 31 Genome wide mapping of Nr5a2 binding sites…………… ……………87 Figure 32 ChIP assay to investigate the binding of Nr5a2 to the Nanog enhancer during the reprogramming of MEFs………………………………………….………88 ... performed the screen by introducing each of these factors in combination with OSKM into human skin fibroblasts Alkaline phosphatase (AP) staining was performed to quantitate the number of human iPSCs ... for reprogramming55,66 Nonetheless, the omission of c-Myc in the cocktail of reprogramming factors results 16 in a marked reduction of efficiency in the generation of iPSCs55,66 Strikingly, N-Myc...

Ngày tải lên: 09/09/2015, 18:56

... 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 ... tri) methylation of lysine of histone H3; H3K27ac, acetylation of lysine 27 of histone H3; H3K27me3, trimethylation of lysine 27 of histone H3; H3K36me3, trimethylation of lysine 36 of histone H3; ... Figure The combination of stem cell models and epigenomics in studies of the role of non-coding mutations in human disease Epigenomic analyses of cells derived through in vitro stem cell differentiation...

Ngày tải lên: 11/08/2014, 12:21

... hinder widespread applications of iPSCs for various applications In addition, in the case of the recombinase-based approach, residual vector sequences are left behind, increasing the risk of insertional ... of 48 iPSC lines established, two contained only a single copy of the polycistronic sequence, indicating that the reprogramming factors are sufficient in single copy for reprogramming The transient ... expression of PB transposase in the two singlecopy cell lines and subsequent subcloning resulted in the removal of the linked reprogramming factor DNA at an efficiency greater than 2%; within these...

Ngày tải lên: 14/08/2014, 21:20

... of LIF in the culture of porcine IPS cells 90 6.4 Importance of doxycycline in the culture media 91 6.5 Differentiation of porcine IPS cells into cardiomyocytes 92 6.6 Use of porcine cells ... fibroblasts to Induced Pluripotent Stem Cells 45 iv 4.10 Culture of Porcine Induced Pluripotent Stem Cells 45 4.10 Characterization of Porcine Induced Pluripotent Stem Cells 46 4.10.1 ... Lin28………… 26 3.4 Cardiac differentiation of Human IPS cell lines 27 3.5 Generation of Porcine Induced Pluripotent Stem Cells 29 3.5.1 Factors used for reprogramming of porcine cells...

Ngày tải lên: 30/09/2015, 10:13

... of pluripotent stem cells Induced pluripotent stem cells attract great interest for their potential in understanding the basics of cell reprogramming, personalized medicine and disease modeling—a ... capability of proliferating indefinitely as undifferentiated cells when cultured in appropriate conditions, and of sustaining a normal karyotype, hESCs may have broad applications for industrial uses; clinical ... antibodies into the blastocyst, thus leaving the ICM cells intact Incubation with guinea pig complement-containing medium then lyses all antibodymarked cells The intact ICM is further rinsed and...

Ngày tải lên: 22/03/2014, 09:20

... selective serotonin reuptake inhibitors In HD, in brains of HD patients and after quinolinic acid striatal lesioning of adult brain the enhanced neurogenesis in the SVZ leads to the migration of neuroblasts ... neurogenesis is increased in the SVZ of brains with HD Data from R6/1 transgenic mouse model of HD are difficult to interpret in the context of adult neurogenesis in HD, as mutated forms of huntingtin affect ... neuronal cells in the adult brain originate from residual stem cells The existence of stem cells in the adult brain suggests that it has the potential for self-repair and that newborn neuronal cells...

Ngày tải lên: 08/08/2014, 17:20

... laboratories using multiple reprogramming methods Coding point mutations were found in all 22 lines, with an average of five protein-coding mutations per line Some of these mutations were present in the ... hallmarks include intracellular accu­ u­ations m l of α-synuclein protein in the form of Lewy bodies and Lewy neurites, and selective loss of dopamin­ rgic (DA) e neurons in the substantia nigra of ... A wide range of gene-targeting techniques, including homologous recombination and the use of zinc-finger nucleases, have been successfully applied to iPSCs [39-41] Adding or removing the AD mutations,...

Ngày tải lên: 11/08/2014, 12:21

... defects in humans [5,6] However, to date, the delivery of MSCs into cartilaginous lesions has neither clinically nor experimentally resulted in sustained regeneration of hyaline cartilage in vivo ... or remained uninfected For comparison, an additional set of untransduced recombinant human protein controls were maintained, which were cultured in the presence of 10 ng/mL TGFβ1 protein, or ... of the double labelling staining solution for 10 minutes at room temperature After staining, aggregates were washed five times with 100 μL of binding buffer, fixed overnight in PBS-buffered 4%...

Ngày tải lên: 09/08/2014, 14:22

... chromatin-binding proteins and modifiers, proteinmodifying factors, and chromatin assembly proteins Importantly, knockdown of Oct4 protein levels is known to cause the loss of co-binding activity of ... expression signature of cancer cells [9] LIN28 Maintains ESC pluripotency by binding and inhibiting the maturation of pro-differentiation let-7 miRNA; LIN28 is also a hiPSC reprogramming factor [74] ... Medicine 2011, 3:68 http://genomemedicine.com/content/3/10/68 Page of 12 Table Dysregulation of transcriptional networks in stem cells and disease Gene/protein Role in ESCs Role in disease c-MYC Involved...

Ngày tải lên: 11/08/2014, 12:21

... Mesenchymal stem cells (MSCs) inhibit the proliferation of CD4 + T cells in vitro by induction of nitric oxide and prostaglandin E2 (PGE2) (a) CD4+ T cells, in the presence of accessory cells, were ... upregulate expression of PD-L1, iNOS, and COX-2 in MSCs, making these molecules candidate mediators of T-cell inhibition The involvement of iNOS and COX-2 in inhibition of T-cell proliferation ... addition of inhibitors of these enzymes - GW274150 and indomethacin [8,36], respectively - to the co-cultures The addition of these inhibitors resulted in the abrogation of the inhibition of Tcell...

Ngày tải lên: 12/08/2014, 11:23

... use of pre-warmed and pre-equilibrated culture media [15, 16] Indeed, minimizing low temperature exposure of human embryos and oocytes is a critical factor in determining the success of clinical ... of ATP in the dynamics of the actin filaments of the cytoskeleton Biol Chem 2006;387(4):401-6 Lopez LA, Bertini F The in vivo effect of low body temperatures on the degree of polymerization of ... layer of mitomycin C-inactivated murine embryonic fibroblast feeder (MEF) cells [7, 8] These were harvested from CF1 inbred mouse strain purchased from Charles River Laboratories Inc (Wilmington,...

Ngày tải lên: 31/10/2012, 16:57

... could maintain a small accessible region in the globin locus during the differentiation of hematopoietic stem cells to erythroid cells Transcription of HS3 could be important in maintaining this ... presence of residual undifferentiated cells We next analyzed the interaction of RNA Pol II and TATA binding protein (TBP) as well as the appearance of modified histones within the globin locus during ... would be to Globin locus activation during differentiation determine the mechanisms involved in this process at the level of individual gene loci In this study, we investigate chromatin structure...

Ngày tải lên: 07/03/2014, 12:20

... [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 directly ... 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 two types of adult cells HMVEC and HUVEC Differences in the ... These included 38 miRNA upregulated in hES cells, 31 upregulated in EB cells, and 35 upregulated in adult cells (Figure 2) The 20 miRNAs most highly expressed in hES cells, EB, and adult cells...

Ngày tải lên: 18/06/2014, 15:20

... are linked to three cellular pathways including p53, phosphatidyl inositol (PI3) kinase, and apoptosis The 14-3-3 proteins are a family of multifunctional phosphoserine/phosphothreonine binding ... factor that is involved in embryonic development and regulation of gene expression in differentiated tissues [37] FOXA1 is involved in regulating apoptosis by inhibiting expression of the anti-apoptotic ... forms of cancers [10] Phosphorylation and dephosphorylation of specific amino acid residues, including serine, tyrosine and threonine, by specific kinases and phosphatases is a major form of post-translational...

Ngày tải lên: 18/06/2014, 19:20

... then incubated with a newly filtered Oil Red O staining solution After staining, the cells were rinsed with distilled water, and ml of isopropyl alcohol was added to the stained dish Aliquots of ... signal intensity values derived from the ROIs were plotted against time as time-intensity curve (TIC) using the Gyroview software system (Philips) The baseline value (SIbase) of the SI in a TIC ... [12] MSCs were cultured in basal medium containing Dulbecco's modified Eagle's medium (DMEM) with 10% fetal bovine serum, 1% mixture of penicillin, streptomycin and neomycin (Invitrogen Corporation,...

Ngày tải lên: 20/06/2014, 00:20

... are linked to three cellular pathways including p53, phosphatidyl inositol (PI3) kinase, and apoptosis The 14-3-3 proteins are a family of multifunctional phosphoserine/phosphothreonine binding ... factor that is involved in embryonic development and regulation of gene expression in differentiated tissues [37] FOXA1 is involved in regulating apoptosis by inhibiting expression of the anti-apoptotic ... forms of cancers [10] Phosphorylation and dephosphorylation of specific amino acid residues, including serine, tyrosine and threonine, by specific kinases and phosphatases is a major form of post-translational...

Ngày tải lên: 20/06/2014, 04:20

... maintains the stem state of MSCs from a variety of species by prolonging their viability in culture [16], sometimes in a cell-autonomous fashion [17] This is reminiscent of the maintenance of ... cells, increased in vivo reconstitution of hematopoietic lineages, and Wnt3a-specific maintenance of skin and intestinal stem cell populations [18] Because stem cells may share signaling mechanisms ... regulate MSC maintenance [19], as they in the selfrenewal of hematopoietic, neural, intestinal, and skin stem cells [18] Wnt3a treatment increases adult MSC proliferation while inhibiting their osteogenic...

Ngày tải lên: 09/08/2014, 10:20

... Expression of BMP-2 and chordinstem cells during osteogenic differentiation of human mesenchymal stem cells (a) Temporal progression of bone morphogenetic protein (BMP)-2 and chordin expression during ... differentiation of MSCs [8,11,12] and promote fracture healing in animal models [13-15] Recombinant human BMP-2 and BMP-7 are used clinically in spinal fusion and the healing of tibial fractures To obtain ... to increased deposition of mineral at late time points The suppression of chordin led to an increase in the bioavailability of endogenously produced BMP-2 to drive the differentiation of Page of...

Ngày tải lên: 09/08/2014, 10:23

... expression of insulin and proinsulin in stage IV cells Quantification of IHC staining shows FN and LAM increases the percentage of (B) proinsulin, and (C) insulin positive cells (mean ± S.D.; ** indicates ... Immunohistochemistry (IHC) in stage IV cells further revealed adding ECM increased the percentage of proinsulin and insulin expressing cells with the maximum effect seen in cells treated with LAM ... widespread interest in finding alternative sources of b -cells for tissue replacement strategies in diabetes MSCs have been used for cell-based therapy in regenerative medicine and tissue engineering In...

Ngày tải lên: 10/08/2014, 05:21